| [construction of probes for hybridization with hepatitis a virus rna based on phage m13]. | three fragments of a dna copy complementary to tre hepatitis a virus rna have been isolated from the phav-vpi-22 plasmid and cloned in m 13 mp 8 vector. the 140, 250 and 390 b. p. fragments corresponded to the viral genome fragment coding for vp1 protein. single-stranded dnas of the hybrid phages with 250 b. p. and 390 b. p. inserts have been used for radioactive probe synthesis. the specificity of the probes for viral rna was confirmed in hybridization with hepatitis a virus rna purified from t ... | 1988 | 2848193 |
| novel eukaryotic expression vectors which permit single-stranded replication in escherichia coli and in vitro translational analysis of cloned genes. | the ability to express cloned genes transiently is an important technique in the study of eukaryotic gene expression. numerous useful expression vectors have been constructed for this purpose although many of them share several common drawbacks. in this paper i describe the construction and characterization of novel expression vectors psvkii and psvkiii which have 13 and 8 unique restriction sites, respectively, suitable for cloning genes. these vectors have phage m13 ori, which permits them to ... | 1988 | 2850970 |
| convenient transfer of lacz-gene fusions to phage m13 by in vivo recombination and their use for nucleotide sequencing. | we describe a method that facilitates the sequencing of lacz fusion joints based on in vivo subcloning onto phage m13. the method is useful for lacz fusions that are isolated with the transposable lambda placmu phage into plasmids carrying the pbr322 bla gene. in vivo cloning of lacz fusions is accomplished by recombination with two m13 phages carrying 5' or 3' segments of the bla gene, adjacent but differing in orientation to lacz'. the presented method allows rapid sequencing of many fusion jo ... | 1988 | 2850975 |
| insertional restoration of beta-galactosidase alpha-complementation (white-to-blue colony screening) facilitates assembly of synthetic genes. | the assembly of synthetic genes from oligodeoxynucleotides can be an inefficient process. upon ligation of a synthetic assembly into a plasmid vector and transformation of an escherichia coli host, it is often found that only a minor fraction of the putative recombinant plasmids contains synthetic sequences. moreover, the synthetic sequences cloned are often altered versions of those originally designed. we have designed a biological test to detect those plasmids that contain synthetic sequences ... | 1988 | 2853687 |
| resonance raman spectroscopy of complexes of the helix destabilizing proteins gp32 and gp5 with poly(ra) and poly(da). | the bacteriophage t4 helix destabilizing protein (hdp) gp32 and its complexes with poly(ra) and poly(da) were studied with ultra-violet resonant raman spectroscopy. the uv-resonant raman (uv-rr) spectrum of the complex of gp5, the coat protein of bacteriophage m13, with poly(da) was also measured and is compared with the spectrum of the gp 32/poly(da) complex. the excitation wavelength was 245.1 nm. this is on the far uv-side of the first absorption bands of adenine and near a "window" in the pr ... | 1988 | 2856034 |
| molecular characteristics of prion rods purified from scrapie-infected hamster brains. | purification of scrapie prions from hamster brains has demonstrated that the infectious particles contain one major protein, prp 27-30. this protein, which is required for and inseparable from scrapie infectivity, polymerizes into heterogeneous rod-shaped particles measuring 10-20 nm in diameter and 100-200 nm in length. we attempted to identify the minimal infectious unit by disrupting aggregates of the rods. prolonged sonication resulted in progressive fragmentation of the rods into spherical ... | 1986 | 2872252 |
| detection of human dna polymorphisms with a simplified denaturing gradient gel electrophoresis technique. | single base pair differences between otherwise identical dna molecules can result in altered melting behavior detectable by denaturing gradient gel electrophoresis. we have developed a simplified procedure for using denaturing gradient gel electrophoresis to detect base pair changes in genomic dna. genomic dna is digested with restriction enzymes and hybridized in solution to labeled single-stranded probe dna. the excess probe is then hybridized to complementary phage m13 template dna, and the r ... | 1987 | 2883652 |
| restriction enzyme analysis of the mitochondrial genome in mitochondrial myopathy. | the mitochondrial myopathies are a heterogeneous group of disorders some of which may be caused by mutations in the mitochondrial genome. mitochondrial dna from 10 patients with mitochondrial myopathy and their mothers was analysed using five restriction enzymes and 11 mitochondrial probes in bacteriophage m13. no abnormalities were found in seven out of the 10 patients. polymorphisms which have not previously been reported were detected in three patients and two of their mothers. these results ... | 1988 | 2903249 |
| singlet molecular oxygen causes loss of biological activity in plasmid and bacteriophage dna and induces single-strand breaks. | damage of plasmid and bacteriophage dna inflicted by singlet molecular oxygen (1o2) includes loss of the biological activity measured as transforming capacity in e. coli and single-strand break formation. three different sources of 1o2 were employed: (i) photosensitization with rose bengal immobilized on a glass plate physically separated from the solution; (ii) thermal decomposition of the water-soluble endoperoxide 3,3'-(1,4-naphthylidene) dipropionate (ndpo2); and (iii) microwave discharge. l ... | 1989 | 2920171 |
| (dt-dc)n and (dg-da)n tracts arrest single stranded dna replication in vitro. | previous in vivo studies have indicated that (dt-dc)n.(dg-da)n tracts (referred to here as (tc)n.(ga)n), which are widely dispersed in vertebrate genomes, may serve as pause or arrest signals for dna replication and amplification. to determine whether these repeat elements act as stop signals for dna replication in vitro, single stranded dnas including (tc)n or (ga)n tracts of various lengths, were prepared by cloning such tracts into phage m13 vectors, and were replicated with the klenow fragme ... | 1989 | 2922274 |
| replication of uv-irradiated single-stranded dna by dna polymerase iii holoenzyme of escherichia coli: evidence for bypass of pyrimidine photodimers. | replication of uv-irradiated circular single-stranded phage m13 dna by escherichia coli rna polymerase (ec 2.7.7.6) and dna polymerase iii holoenzyme (ec 2.7.7.7) in the presence of single-stranded dna binding protein yielded full-length as well as partially replicated products. a similar result was obtained with phage g4 dna primed with e. coli dna primase, and phage phi x174 dna primed with a synthetic oligonucleotide. the fraction of full-length dna was several orders of magnitude higher than ... | 1986 | 2941756 |
| effects of escherichia coli ssb protein on the single-stranded dna-dependent atpase activity of escherichia coli reca protein. evidence that ssb protein facilitates the binding of reca protein to regions of secondary structure within single-stranded dna. | the effect that escherichia coli single-stranded dna binding (ssb) protein has on the single-stranded dna-dependent atpase activity of reca protein is shown to depend upon a number of variables such as order of addition, magnesium concentration, temperature and the type of single-stranded dna substrate used. when ssb protein is added to the dna solution prior to the addition of reca protein, a significant inhibition of atpase activity is observed. also, when ssb protein is added after the format ... | 1987 | 2953903 |
| the occurrence of 2'-5' oligoadenylates in escherichia coli. | the use of a highly specific radioimmunoassay and of hplc permitted us to confirm the occurrence of 2'-5' oligoadenylates [p chi (a2'p5')na] in several strains of escherichia coli. cellular concentrations of 2'-5' oligoadenylates ranged from 50 nm to 300 nm. the mixture of 2'-5' oligoadenylates consisted primarily of pppa2'p5'a, pa2'p5'a,a2'p5'ap and a2'p5'a under normal conditions of growth. none of them activated rnase l. infection of the bacteria with the single-stranded dna phage m13 or indu ... | 1987 | 2960522 |
| high-level production of hepatitis b viral x protein in escherichia coli using gene ii promoter of bacteriophage m13. | region x is one of the four open reading frames (orfs) of hepatitis b virus (hbv) and encodes a polypeptide of 154 amino acids (aa). a 584-bp bamhi-bglii fragment of the hbv dna containing the major part of orf x which encodes 145 aa was inserted into the bglii site within the gene ii of bacteriophage m13. the insertion resulted in an in-phase gene ii-x fused protein of 174 aa under the control of the gene ii promoter. cells harboring plasmids (pml alpha x.59 and pmlx.12d) derived from the above ... | 1988 | 2966757 |
| genome interactions which influence dna palindrome mediated instability and inviability in escherichia coli. | the interaction of three factors determine the detrimental effect of a palindromic dna sequence in escherichia coli cells. the first is the nature of the palindrome (its length, extent of central asymmetry and perhaps its base sequence), the second is the genotype of the host cell and the third is the replicon within which it is located. in this paper we extend the genetic and physical characterization of lambda bacteriophages carrying a palindrome of approximately 560 base pairs. we also show t ... | 1987 | 2972734 |
| improved m13 phage cloning vectors and host strains: nucleotide sequences of the m13mp18 and puc19 vectors. | three kinds of improvements have been introduced into the m13-based cloning systems. (1) new escherichia coli host strains have been constructed for the e. coli bacteriophage m13 and the high-copy-number puc-plasmid cloning vectors. mutations introduced into these strains improve cloning of unmodified dna and of repetitive sequences. a new suppressorless strain facilitates the cloning of selected recombinants. (2) the complete nucleotide sequences of the m13mp and puc vectors have been compiled ... | 1985 | 2985470 |
| cloning and dna sequence of a plasmid-determined citrate utilization system in escherichia coli. | the citrate utilization determinant from a large 200-kilobase (kb) naturally occurring plasmid was previously cloned into the psti site of plasmid vector pbr325 creating the cit+ tetracycline resistance plasmid pwr61 (15 kb). tn5 insertion mutagenesis analysis of plasmid pwr61 limited the segment responsible for citrate utilization to a 4.8-kb region bordered by ecori and psti restriction nuclease sites. the 4.8-kb fragment was cloned into phage m13, and the dna sequence was determined by the di ... | 1985 | 2999088 |
| screening for thermostable mutant of kanamycin nucleotidyltransferase by the use of a transformation system for a thermophile, bacillus stearothermophilus. | a structural gene of kanamycin nucleotidyltransferase cloned into a single-stranded bacteriophage m13 was subjected to mutagenesis with hydroxylamine. having recloned the mutagenized gene of the enzyme in a vector plasmid ptb922, the recombinant plasmid was used to transform bacillus stearothermophilus with a purpose of screening for the more thermostable enzyme than the wild type. out of greater than 8 x 10(3) transformants, 12 clones that were suspected to harbor the mutant gene encoding the m ... | 1985 | 2999110 |
| a complete library of point substitution mutations in the glucocorticoid response element of mouse mammary tumor virus. | the glucocorticoid response element (gre) of mouse mammary tumor virus (mmtv) was chemically synthesized as two complementary dna strands bearing cohesive termini. during automated synthesis, random mutations were introduced into the dna by "doping" each of the four nucleoside phosphoramidites (a, g, c, and t) with a low level of the other three. these preparations were annealed and cloned into an m13 phage vector to produce a library of gre mutants. mutations within the synthesized region were ... | 1986 | 3003746 |
| illegitimate recombination at the replication origin of bacteriophage m13. | hybrids composed of phage m13 and plasmid phv33 were used to study the formation of deletions in escherichia coli. eighty to ninety percent of the deletion endpoints were at the position of the nick introduced into the m13 replication origin by the phage gene ii protein. this suggests the existence of a novel mechanism of illegitimate recombination. | 1986 | 3010295 |
| dna alterations photosensitized by tetracycline and some of its derivatives. | bacteriophage m13 mp10 dna were irradiated with near-uv light in the presence of tetracycline derivatives and primed with synthetic oligonucleotide to be used for dna synthesis using escherichia coli dna polymerase. chain terminations were observed by denaturing polyacrylamide gel electrophoresis and mapped precisely. all the synthesis stops occurred before or at the level of guanine residues, showing that the photoreaction mediated by tetracycline derivatives led to a preferential alteration of ... | 1986 | 3011916 |
| hydrolysis by restriction endonucleases at their dna recognition sequences substituted with mismatched base pairs. | restriction endonucleases were tested for their ability to catalyze the cleavage of mismatch-containing recognition sites in dna. these mismatched base pairs were t.g, u.g, or a.c in covalently closed, circular heteroduplexes prepared by in vitro extension of chemically synthesized oligonucleotide primers annealed to a bacteriophage m13-derived viral dna. none of the restriction enzymes was able to completely cleave the mismatch-containing recognition sites under standard conditions. however, th ... | 1986 | 3012472 |
| nucleotide sequence and deduced amino acid sequence of escherichia coli pyruvate oxidase, a lipid-activated flavoprotein. | the entire nucleotide sequence of the poxb (pyruvate oxidase) gene of escherichia coli k-12 has been determined by the dideoxynucleotide (sanger) sequencing of fragments of the gene cloned into a phage m13 vector. the gene is 1716 nucleotides in length and has an open reading frame which encodes a protein of mr 62,018. this open reading frame was shown to encode pyruvate oxidase by alignment of the amino acid sequences deduced for the amino and carboxy termini and several internal segments of th ... | 1986 | 3016647 |
| site-directed mutagenesis of the binding site for ribosomal protein s8 within 16s ribosomal rna from escherichia coli. | twelve specific alterations have been introduced into the binding site for ribosomal protein s8 in escherichia coli 16s rrna. appropriate rdna segments were first cloned into bacteriophage m13 vectors and subjected to bisulfite and oligonucleotide-directed mutagenesis in vitro. subsequently, the mutagenized sequences were placed within the rrnb operon of plasmid pno1301 and the mutant plasmids were used to transform e. coli recipients. the growth rates of cells containing the mutant plasmids wer ... | 1986 | 3016664 |
| tarantula hemocyanin mrna. in vitro translation, cdna cloning and nucleotide sequence corresponding to subunit e. | following induction of hemopoiesis, poly(a)-rich rna was prepared from the heart of the tarantula, eurypelma californicum, and translated in rabbit reticulocyte lysates. in vitro translation products were immunoprecipitated with antiserum against whole dissociated eurypelma hemocyanin. analysis of the immunoprecipitate by sodium dodecyl sulfate/polyacrylamide gel electrophoresis revealed a set of polypeptides comigrating with authentic eurypelma hemocyanin. the mrna was transcribed into cdna, cl ... | 1986 | 3017715 |
| nucleotide sequence of cdna and derived amino acid sequence of rabbit complement component c3 alpha-chain. | the nucleotide sequence coding for 726 amino acid residues of the alpha-chain of rabbit c3 was determined from a cdna clone. subfragments of the cdna produced by restriction endonucleases were inserted into the bacteriophage m13 and sequenced using the dideoxynucleotide technique. the derived amino acid sequence was compared with those of human and mouse c3, which have been previously reported [by de brujn, m.h.l. and fey, g.h. (1985) proc. natl. acad. sci. usa 82, 708, and westel, r.a. et al. ( ... | 1986 | 3019881 |
| nucleotide sequence of the structural genes for an anion pump. the plasmid-encoded arsenical resistance operon. | the structural genes for the arsenical pump of the conjugative r-factor r773 contained on a hindiii fragment of 4.3 kilobase pairs were cloned into bacteriophage m13. a series of ordered deletions was created using bal31 digestion, and the nucleotide sequence of the operon determined. three open reading frames for genes arsa, arsb, and arsc were found. the arsa gene encodes a hydrophilic protein of 63,169 da with two potential adenylate-binding sites. the arsb gene encodes a potentially membrane ... | 1986 | 3021763 |
| [insertion of the cos-site into dna of phage m13 and its packing in proteins of phage lambda]. | the cos-site of lambda phage from phc79 cosmide is transferred to dna from m13 mp18 phage. the recombinant dna thus obtained (mc18) is efficiently packaged into lambda proteins in vitro. the bamhi-hindiii fragment of pgp588 (a pbr322 derivatives containing fragment of human dna) is subcloned into mc18. although this pgp588 fragment contains numerous alu repeats, no essential rearrangements of the insert were revealed. the efficiency infection by recombinant dna packaged with lambda proteins is a ... | 1986 | 3022756 |
| characterization of the ftsb gene as an allele of the nrdb gene in escherichia coli. | a temperature-sensitive, salt-rescuable ftsb cell division mutant, mft84, was found to be hydroxyurea sensitive on low-salt medium. complementation studies with plasmids and a marker rescue study with bacteriophage m13 nrd indicated that ftsb is an allele of nrdb and that the mutation occurs in the region corresponding to nucleotides 6729 to 7032 of the nrdb gene. enzymatic characterization demonstrated that the b2 subunit of ribonucleoside-diphosphate reductase encoded by ftsb was responsible f ... | 1987 | 3025167 |
| a simple and rapid nucleotide sequencing strategy and its application in analyzing a rice histone 3 gene. | an improved rapid method for sequencing a target dna is described. a new plasmid, paa-pz1, which contains the origin of replication from phage m13 and a portion of the tn9 transposon was constructed. a long fragment of target dna cloned into this vector is progressively shortened in vivo from one end by transposon-mediated deletions. the plasmids carrying different lengths of target dna are then made into single-stranded dna in the same host upon infection with an m13 phage and their sequence is ... | 1986 | 3026911 |
| expression of a trna gene in the context of the lacz mrna. | fusions of the gene for tyrosine suppressor trna, tyrt(sup3), and the lacz gene of escherichia coli were constructed such that the trna gene could be expressed from either its own promoter or that of the lac operon. these chimeras, carried on phage m13 vectors, were tested for the expression of the trna in e. coli. the trna gene was expressed on the order of 10-fold more weakly from the lac promoter than from its own promoter. to examine whether pausing or premature termination of transcription ... | 1987 | 3027034 |
| analysis of the role of the cysteine 171 residue in the activity of herpes simplex virus type 1 thymidine kinase by oligonucleotide-directed mutagenesis. | the thymidine kinase (tk) gene from herpes simplex virus type 1 strain sc16 was cloned into bacteriophage m13 mp8 so that functional hsv-1 tk was expressed in bacteria infected with the recombinant bacteriophage, m13/tk. oligonucleotide site-directed mutagenesis was then employed to introduce single nucleotide changes into the tk gene in m13/tk in order to alter the codon for cysteine 171 in the wild-type enzyme to a codon specifying either serine or glycine. analysis of the mutant enzymes in ba ... | 1987 | 3027247 |
| coliphage m13 cloning system and ddxtp chain-termination method for dna sequencing. | two dna fragments of cytochrome b gene in yeast mitochondrial dna were sequenced by messing's m13 cloning system and sanger's ddxtp chain-termination method. m13mp8 and m13mp9 serve as vectors for insert fragment. the recipient strain is e. coli jm103 for transfection. when the ratio of insert/vector was 3:1, high frequencies of recombination and positive recombination were obtained. the two fragments, which have 575 bp and 709 bp, were sequenced. to read more bases, the ratio of ddxtp/dxtp must ... | 1986 | 3027889 |
| [variants of phage m13 dna containing a fragment of the beta-galactosidase gene--a convenient mutation system for the study of oligonucleotide-directed mutagenesis]. | a model system is developed to test oligonucleotide-directed mutations: t----c transition, t and c deletions (delta t and delta c), c insertion, double mutations (a----g, delta t), (t----c, a----g), and large oligonucleotide deletions (36 or 44 nucleotides). the system includes 9 variants of the phage m13 dna carrying fragment of beta-galactosidase gene, and oligodeoxyribonucleotides partially noncomplementary to dna sequence of this gene. six variants are obtained by the site-localized mutagene ... | 1986 | 3028430 |
| determination of size and orientation of dna fragments cloned in phage m13 by s1 nuclease mapping. | | 1987 | 3029677 |
| differential gene expression during the amoebal-plasmodial transition in physarum. | we have prepared cdna libraries for amoebae and plasmodia of the acellular slime mould, physarum polycephalum. differential screening was used to isolate cell-type-specific cdna clones (in bacteriophage m13) and both libraries yielded approximately 5% of such sequences. the amoebal- and plasmodial-specific clones were used to assay changes in transcription during the amoebal-plasmodial transition. the results obtained substantiate the view that the switch from amoebal to plasmodial characteristi ... | 1987 | 3029710 |
| recombinant forms of m13 procoat with an ompa leader sequence or a large carboxy-terminal extension retain their independence of secy function. | the assembly of phage m13 procoat protein into the plasma membrane of escherichia coli is independent of the secy protein. to test whether this is caused by the unusually small size of procoat, we fused dna encoding 103 amino acids to the carboxy-terminal end of the procoat gene. the resulting fusion protein, which attains the same membrane-spanning conformation as mature coat protein, still does not require the secy function for membrane assembly. to determine whether the leader sequence govern ... | 1987 | 3034592 |
| dna mismatch-repair in escherichia coli counteracting the hydrolytic deamination of 5-methyl-cytosine residues. | derivatives of phage m13 were constructed and used for the in vitro preparation of heteroduplex dna molecules containing base/base mismatches that mimick dna lesions caused by hydrolytic deamination of 5-mec residues in escherichia coli dna (i.e. they carry a t/g mismatch in the special sequence context provided by the recognition site -cca/tgg-of the dcm-methyltransferase). upon introduction of these heteroduplex dnas into cacl2-treated e. coli cells, the mismatches are efficiently repaired wit ... | 1987 | 3038536 |
| effects of temperature-sensitive variants of the bacillus subtilis dnab gene on the replication of a low-copy-number plasmid. | the dnab gene of bacillus subtilis is involved in the initiation of dna replication and also in the binding of the chromosomal origin to the bacterial membrane. we studied the effect of temperature-sensitive dnab mutants (dnab1 and dnab19) on the replication and on the dna-membrane binding of the plasmid pkw1, which was derived from the low-copy-number plasmid pbs2. in the dnab19 mutant, pkw1 was not able to replicate at the restrictive temperature. in the dnab1 mutant, however, the dimeric form ... | 1987 | 3040678 |
| structure and dynamics of a detergent-solubilized membrane protein: measurement of amide hydrogen exchange rates in m13 coat protein by 1h nmr spectroscopy. | the coat protein of bacteriophage m13 is inserted into the inner membrane of escherichia coli where it exists as an integral membrane protein during the reproductive cycle of the phage. the protein sequence consists of a highly hydrophobic 19-residue central segment flanked by an acidic 20-residue n-terminus and a basic 11-residue c-terminus. we have measured backbone amide hydrogen exchange of the protein solubilized in perdeuteriated sodium dodecyl sulfate using 1h nuclear magnetic resonance ( ... | 1988 | 3042022 |
| nucleotide sequence analysis and overexpression of the gene encoding a type iii chloramphenicol acetyltransferase. | the gene catiii, encoding a type iii enterobacterial chloramphenicol acetyltransferase, was cloned from the transmissible plasmid r387 into pbr322 and bacteriophage m13 mp8. nucleotide sequence analysis of 1160 bp of dna identified an open reading frame encoding a protein of 213 amino acid residues and a calculated molecular mass of 24965 da. the predicted n-terminal sequence is identical with that determined by edman degradation of chloramphenicol acetyltransferase purified from escherichia col ... | 1988 | 3048245 |
| [genomic fingerprinting of microorganisms: its use as a hybridization probe of phage m13 dna]. | hypervariable nucleotide sequences detected by hybridization with the phage m13 dna probe were found in the chromosomal dnas of certain pathogenic microbial species. dna fingerprinting, based on hybridization of m13-probe with hypervariable chromosomal dna sequences, opens new approaches to epidemiological analysis, epidemiological prognosis, taxonomy, and other theoretical and applied fields of bacteriology. | 1988 | 3053332 |
| a novel replicon occurring naturally in escherichia coli is a phage-plasmid hybrid. | a novel dna replicon in escherichia coli was identified. it is the smallest natural isolate (1282 bp) found so far. in the presence of phage m13 it grows as a filamentous single-stranded dna phage. contrary to previously identified mini-phages this replicon displays sequence homology only to parts of the m13 viral and complementary strand origin. in the absence of m13 this dna replicates autonomously. the only gene (arp) of the replicon encodes a 32-kd protein, which is essential for autonomous ... | 1988 | 3061812 |
| [a simple system of cloning in phage m13 and dna sequencing with terminators]. | a compilation of techniques for dna cloning in filamentous phage m13 based vectors for a novice in cloning is presented. it does not require either specialized microbiological facilities, or any specific knowledge in escherichia coli genetics. the cloning strategy uses only blunt-end ligation into a vector that has been prepared once for several hundred experiments. the first part describes the isolation, preparation and checking of a blunt-ended m13 vector (with m13 mp series vectors as an exam ... | 1988 | 3065614 |
| [expression in escherichia coli of dna coding for human tumor necrosis factor]. | the variants of expression in escherichia coli of artificial dna coding for human tumor necrosis factor, an important immune modulator with selective cytotoxic action on a number of transformed cell lines have been described. the dna was placed under control of either phage m13 promoter of gene for main coat protein or tandem of pair of e. coli tryptophane promoters. it has been shown that e. coli cells harbouring plasmids described with artificial tnf gene provide good level of protein biosynth ... | 1988 | 3071369 |
| secretion and export of igf-1 in escherichia coli strain jm101. | the processing of lamb-igf-1 fusion protein and the export of processed igf-1 (insulin-like growth-factor-1) into the growth medium was examined in the escherichia coli host strain, jm101. several strain or plasmid modifications were tried to increase export of periplasmic (processed) igf-1 into the growth medium of jm101. these included: (1) use of a lon null mutant strain to increase accumulation levels of unprocessed lamb-igf-1 fusion protein; (2) use of an alternative drug resistance marker ... | 1988 | 3071740 |
| specificity of ionizing radiation-induced mutagenesis in the lac region of single-stranded phage m13 mp10 dna. | m13 mp10 single-stranded phage dna was irradiated with 60 co gamma-rays, and transfected into escherichia coli. one hundred and sixteen mutant clones having lesions in the lac insert were selected, and mutational sites were examined by dna sequence analysis. fourteen out of the 15 nucleotide changes thus detected were base substitutions, and the rest was a base addition. transitions and transversions were almost equal in number. mutational events were observed at cytosine residues more frequentl ... | 1986 | 3088546 |
| purified scrapie prions resist inactivation by uv irradiation. | the development of effective purification protocols has permitted evaluation of the resistance of isolated scrapie prions to inactivation by uv irradiation at 254 nm. prions were irradiated on ice with doses of uv light ranging up to 120,000 j/m2. uv dosimetry experiments, performed with saccharomyces cerevisiae plasmid dna or eucaryotic cells, indicated that under these experimental conditions an incident uv dose of 10 j/m2 formed 2 thymine dimers per 5.1 x 10(6) daltons of eucaryotic cell dna. ... | 1987 | 3097336 |
| purified scrapie prions resist inactivation by procedures that hydrolyze, modify, or shear nucleic acids. | prions were purified from scrapie-infected hamster brains and incubated for 24 hr at 65 degrees with 2 mm zn2+ or 5 mm mg2+; no loss of infectivity was observed. bacteriophage m13, tobacco mosaic virus (tmv), potato virus x, and potato spindle tuber viroid were all inactivated by divalent metal ions under these conditions. prions also resisted inactivation by prolonged digestions with dnase i, rnases a and t1, and micrococcal nuclease. prions were resistant to psoralen photoadduct formation usin ... | 1987 | 3114950 |
| cloning and sequence analysis of cytadhesin p1 gene from mycoplasma pneumoniae. | mycoplasma pneumoniae cytadhesin p1 was purified by monoclonal antibody affinity chromatography followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. the n-terminal 18-amino-acid sequence of p1 was determined and used to design two synthetic oligonucleotides, a 14-mer corresponding to amino acids 1 to 5 and an 18-mer corresponding to amino acids 7 to 12. these oligonucleotides served as hybridization probes for the identification of the p1 gene by southern blot analy ... | 1987 | 3119495 |
| preferential uptake of restriction fragments from a gonococcal cryptic plasmid by competent neisseria gonorrhoeae. | factors involved in the specificity of dna uptake by competent neisseria gonorrhoeae were examined. host-controlled modification did not affect uptake. certain restriction fragments of the 4.2 kb gonococcal cryptic plasmid pfa1 and of the replicative form of the bacteriophage m13 were taken up in preference to others, independent of differences in fragment size. a 600 bp fragment from the 4.2 kb plasmid was cloned into ples2, a gonococcal-escherichia coli shuttle vector; the 600 bp fragment was ... | 1988 | 3141569 |
| effects of sos and mucab functions on reactivation and mutagenesis of m13 replicative form dna bearing bulky lesions. | we have previously determined the specificity of -1 frameshifts induced by aflatoxin-b1-2,3-dichloride (afb1c12) in phage m13 double-strand replicative form (rf) dna. the system consists of: (i) in vitro adduction of rf dna of bk8, a lacz + 1 frameshift derivative of phage m13mp8; (ii) transfection into unirradiated or uv-irradiated bacterial host cells; (iii) scoring and sequencing of revertants (i.e., -1 frameshifts). the previous data had shown that induction of sos functions enhanced mutagen ... | 1988 | 3141805 |
| repetitive dna sequences as probes for mycobacterium tuberculosis. | three cloned segments of mycobacterium tuberculosis dna which are promising as clinical probes were identified. an mboi digest of dna from a clinical isolate of m. tuberculosis was cloned into bacteriophage m13. to identify recombinants specific for the m. tuberculosis complex, plaque lifts were hybridized with m. bovis and m. kansasii dna. recombinants which selectively hybridized with m. bovis dna were characterized by probing slot blots and restriction digests of dna from various mycobacteria ... | 1988 | 3148630 |
| chemical and biological studies of the major dna adduct of cis-diamminedichloroplatinum(ii), cis-[pt(nh3)2(d(gpg]], built into a specific site in a viral genome. | a duplex escherichia coli bacteriophage m13 genome was constructed containing a single cis-[pt(nh3)2(d(gpg]] intrastrand cross-link, the major dna adduct of the anticancer drug cis-diamminedichloroplatinum(ii). the duplex dodecamer d(agaaggcctaga).d(tctaggccttct) was ligated into the hincii site of m13mp18 to produce an insertion mutant containing a unique stui restriction enzyme cleavage site. a genome with a 12-base gap in the minus strand was created by hybridizing hincii-linearized m13mp18 d ... | 1988 | 3166983 |
| deuterium nuclear magnetic resonance investigation of bacteriophage m13 coat protein in dimyristoylphosphatidylcholine liposomes using palmitic acid as a probe. | the effect of incorporation of various amounts of m13 bacteriophage coat protein on the bilayer order and acyl chain motion in dimyristoylphosphatidylcholine (dmpc) liposomes has been investigated using deuterium nmr of specifically deuterated palmitic acid as a bilayer probe, phosphorus nmr and additional spin-label electron spin resonance (esr). the secondary structure of the m13 coat protein in these bilayers was determined from circular dichroism spectra. phosphorus nmr spectra of the mixed ... | 1988 | 3179303 |
| [complementary addressed modification of single- and double-stranded dna by alkylating derivatives of oligonucleotides isolated by partial dna fragmentation]. | reagents for complementary addressed modification of nucleic acids are proposed to be synthesized on the base of oligonucleotides obtained by partial chemical fragmentation of dna. the alkylating 4-(n-2 chlorethyl-n-methylamino) benzyl-5'-phosphamide derivatives of 5'-[32p]-labelled oligonucleotides obtained from single and double-stranded dna cloned in bacteriophage m13 mp9 have been synthesized. the alkylated derivatives of oligonucleotides selectively modify the complementary tracts of single ... | 1988 | 3193969 |
| characterization of a sequence of human t cell leukemia virus type i from a patient with chronic progressive myelopathy. | dna from peripheral blood mononuclear cells of individuals with chronic progressive myelopathy (cpm) were extensively analyzed for the presence of human t cell leukemia virus (htlv) type i-like sequences by using the polymerase chain reaction. the dna samples were amplified with oligonucleotides from three separate regions of htlv viral genomes. a portion of the amplified viral genome from a representative patient was sequenced after molecular cloning into bacteriophage m13. sequence data indica ... | 1988 | 3198935 |
| [complementary-addressed elimination of e1a sequence of simian adenovirus oncogene sa7 from circular single-stranded dna of recombinant phage m13]. | the g fragment of the simian adenovirus sa7 oncogene corresponding to e1a region was cloned into m13mp8 and m13mp9 phages. single-stranded dnas of the recombinant phages thus obtained (mp8g and mp9g) partially digested with dnase ii were used to synthesize polyalkylating derivatives capable of specific hybridisation and subsequent alkylation of complementary g sequences of corresponding phage dnas. after incubation of complementary alkylated dna in the presence of lysine, the preselected region ... | 1988 | 3219135 |
| [effectiveness of distal gene translation in polycistrons depends upon the arrangement of regulatory signals on a template]. | the role of the translational terminator and initiator signals arrangement for two adjacent genes in polycistronic mrna has been studied. semisynthetic beta-galactosidase gene (lacz) of e. coli and fragment of phage m13 dna (with promoter pviii, gene ix, and part of gene viii) were used for constructing of the ix-viii-lacz artificial polycistronic operon. cloning of the constructs into pbr322 vector resulted in a number of plz381n plasmids differing by the mutual arrangement of gene viii transla ... | 1988 | 3233097 |
| cloning and determination of the nucleotide sequence of the mn-containing superoxide dismutase gene from halobacterium halobium. | a group of synthetic 17-mer oligodeoxynucleotides (oligos) was constructed to correspond to a sequence of amino acids situated near the n terminus of the manganese-containing superoxide dismutase (mn-sod) purified from the halophilic bacterium, halobacterium halobium. a cosmid library of a sau3ai partial digest of halobium dna, cloned into the bamhi site of phc79, was probed with the radiolabeled oligos. cosmid dna was purified from the clone that showed hybridization at the highest stringency. ... | 1988 | 3240866 |
| sequence analysis of a processed gene coding for mouse ribosomal protein l32. | the nucleotide sequence of a mouse ribosomal protein gene, identified by hybridization with the gene encoding the drosophila ribosomal (r-) protein 49, was determined by cloning in the phage m13 and dideoxy sequencing. the mouse gene, l32', is a member of the multigene family encoding mammalian r-protein l32. l32' is a processed gene that could encode a 135 amino acid protein similar to that of mouse l32 and drosophila r-protein 49. | 1988 | 3246356 |
| dna-binding properties of gene-5 protein encoded by bacteriophage m13. 2. further characterization of the different binding modes for poly- and oligodeoxynucleic acids. | the binding of gene-5 protein, encoded by bacteriophage m13, to oligodeoxynucleic acids was studied by means of fluorescence binding experiments, fluorescence depolarization measurements and irreversible dissociation kinetics of the protein.nucleotide complexes with salt. the binding properties thus obtained are compared with those of the binding to polynucleotides, especially at very low salt concentration. it appears that the binding to oligonucleotides is always characterized by a stoichiomet ... | 1988 | 3262511 |
| sequence-specific chemical modification of a hybrid bacteriophage m13 single-stranded dna by alkylating oligonucleotide derivatives. | alkylating oligonucleotide derivatives react with the complementary sequences in hybrid m13mp7 bacteriophage single-stranded dna and destroy the infecting ability of the dna. the reagents do not damage m13mp9 single-stranded dna lacking the target nucleotide sequence. | 1988 | 3282926 |
| [genomic fingerprints of organisms from different taxonomic groups: the use of phage m13 dna as a hybridization probe]. | hypervariable polymorphic patterns were detected using wild-type m13 dna as a probe in genomic dnas of very different organisms ranging from procaryotes and lower eucaryotes to upper plants and animals, including human beings. due to somatic stability of highly polymorphic patterns and their discrete inheritance, individual-specific restriction pattern analysis ("dna fingerprinting") with this test probe was found to be useful in applied human genetics, in particular, for identifying paternity a ... | 1988 | 3282990 |
| detection of toxigenic escherichia coli using biotin-labelled dna probes following enzymatic amplification of the heat labile toxin gene. | several types of dna probes labelled with biotin were compared for their sensitivity to detect the heat labile toxin (lt) gene in toxigenic escherichia coli. in addition, a procedure was developed for enzymatically amplifying lt gene sequences in toxigenic e. coli. probes were labelled with biotinylated nucleotides by either nick translation; 3' tailing; primer extension of probe dna cloned into bacteriophage m13; sandwich hybridization; or oligolabelling of isolated dna fragments. a single stra ... | 1988 | 3288864 |
| mutant 16s ribosomal rna: a codon-specific translational suppressor. | we have isolated an unusual codon-specific translational suppressor in escherichia coli. the suppressor resulted from a spontaneous mutation in a chromosomal gene during a selection for suppressors of the auxotrophic nonsense mutation trpa(uga211). the suppressor allows readthrough of uga mutations at two positions in trpa and at two sites in bacteriophage t4. it does not, however, suppress amber (uag) or ochre (uaa) mutations that were tested in both genomes, some of which were at the same posi ... | 1988 | 3288986 |
| effects of the escherichia coli ssb protein on the binding of escherichia coli reca protein to single-stranded dna. demonstration of competitive binding and the lack of a specific protein-protein interaction. | the effect of the escherichia coli single-stranded dna binding (ssb) protein on the stability of complexes of e. coli reca protein with single-stranded dna has been investigated through direct dna binding experiments. the effect of each protein on the binding of the other to single-stranded dna, and the effect of ssb protein on the transfer rate of reca protein from one single-stranded dna molecule to another, were studied. the binding of ssb protein and reca protein to single-stranded phage m13 ... | 1987 | 3295259 |
| sequence comparison of single-stranded dna binding proteins and its structural implications. | the primary sequences were compared among several proteins: gene product 5 protein (gp5) from phage m13; pike from phage ike; gene product 32 protein (gp32) from phage t4; reca, ssb and ssf from escherichia coli. these proteins bind strongly and cooperatively to single-stranded dna with no sequence specificity. gp5 is the smallest in this group and its three-dimensional structure is well-characterized. using the entire sequence of gp5 as a template we searched for the regions in other single-str ... | 1987 | 3295261 |
| the dna sequence specificity of stimulation of dna polymerases by factor d. | the mechanism of enhancement of dna polymerase activity by the murine dna-binding protein factor d was investigated. extension by escherichia coli dna polymerase i and calf thymus dna polymerase-alpha of 5'-32p-labeled oligodeoxynucleotide primers that are complementary to poly(dt) or to bacteriophage m13 dna was measured in the absence or presence of factor d. with 5'-[32p](da)9.poly(dt), factor d enables e. coli polymerase i to fill approximately 15-nucleotide gaps between adjacent primers; wh ... | 1987 | 3298245 |
| high frequencies of short frameshifts in poly-ca/tg tandem repeats borne by bacteriophage m13 in escherichia coli k-12. | slipped-strand mispairing (ssm) may play an major role in repetitive dna sequence evolution by generating large numbers of short frameshift mutations within simple tandem repeats. here we examine the frequency and size spectrum of frameshifts generated within poly-ca/tg sequences inserted into bacteriophage m13 in escherichia coli hosts. the frequency of detectable frameshifts within a 40 bp tract of poly-ca/tg is greater than one percent and increases more than linearly with length, being lower ... | 1987 | 3299269 |
| inhibition of purified escherichia coli leader peptidase by the leader (signal) peptide of bacteriophage m13 procoat. | the leader peptide of bacteriophage m13 procoat inhibited the cleavage of m13 procoat or pre-maltose-binding protein by purified escherichia coli leader peptidase. this finding confirms inferences that the leader is the primary site of enzyme recognition and suggests a rationale for the rapid hydrolysis of leader peptides in vivo. | 1987 | 3301818 |
| homologous recombination intermediates between two duplex dna catalysed by human cell extracts. | using as substrates, 1: the replicative form (rf) of phage m13 mp8 in which the reading frame of the lac z' gene was disrupted by insertion of an octonucleotide, and 2: a restriction fragment one kb long, containing the functional lac z' gene (isolated from wild type m13 mp8), we show that nuclear extracts from human cells (3 lines tested) promote the targeted replacement of the altered sequence by the functional one. following incubation with the extracts, the dna's were introduced in jm 109 ba ... | 1987 | 3302944 |
| backbone dynamics of a model membrane protein: assignment of the carbonyl carbon 13c nmr resonances in detergent-solubilized m13 coat protein. | the major coat protein of the filamentous bacteriophage m13 is a 50-residue amphiphilic polypeptide which is inserted, as an integral membrane-spanning protein, in the inner membrane of the escherichia coli host during infection. 13c was incorporated biosynthetically into a total of 23 of the peptide carbonyls using labeled amino acids (alanine, glycine, lysine, phenylalanine, and proline). the structure and dynamics of carbonyl-labeled m13 coat protein were monitored by 13c nuclear magnetic res ... | 1987 | 3307912 |
| deoxyhexanucleotide containing a vinyl chloride induced dna lesion, 1,n6-ethenoadenine: synthesis, physical characterization, and incorporation into a duplex bacteriophage m13 genome as part of an amber codon. | organic synthesis and recombinant dna techniques have been used to situate a single 1,n6-ethenoadenine (epsilon ade) dna adduct at an amber codon in the genome of an m13mp19 phage derivative. the deoxyhexanucleotide d[gct(epsilon a)gc] was chemically synthesized by the phosphotriester method. mild nonaqueous conditions were employed for deprotection because of the unstable nature of the epsilon ade adduct in aqueous basic milieu. physical studies involving fluorescence, circular dichroism, and 1 ... | 1987 | 3314993 |
| solution hybridization of crosslinkable dna oligonucleotides to bacteriophage m13 dna. effect of secondary structure on hybridization kinetics and equilibria. | several dna oligonucleotides have been photochemically modified with the furocoumarin 4'-hydroxymethyl-4,5',8-trimethylpsoralen (hmt) such that each contained a single hmt furan side monoadduct to thymidine at a unique 5' tpa 3' sequence. when these oligonucleotides were hybridized to their respective complements, the hmt adduct could be driven to form an interstrand crosslink by irradiation of the hybrid with 360 nm light. the ability to crosslink probe-target complexes has allowed us to determ ... | 1987 | 3316669 |
| bacteriophage m13 procoat protein inserts into the plasma membrane as a loop structure. | the major coat protein of bacteriophage m13 is synthesized as a precursor, the procoat, with a typical leader (signal) sequence of 23 residues at its nh2-terminus. a fusion protein that contains the nh2-terminal 141 residues of cytoplasmic ribulokinase and all but the first ten residues of m13 procoat was made. the fusion protein inserts into the plasma membrane of escherichia coli and is processed by leader peptidase to give rise to a leader peptide of 155 residues and the mature coat protein o ... | 1987 | 3317833 |
| time-resolved tryptophan fluorescence anisotropy investigation of bacteriophage m13 coat protein in micelles and mixed bilayers. | coat protein of bacteriophage m13 is examined in micelles and vesicles by time-resolved tryptophan fluorescence and anisotropy decay measurements and circular dichroism experiments. circular dichroism indicates that the coat protein has alpha-helix (60%) and beta-structure (28%) in 700 mm sodium dodecyl sulfate micelles and predominantly beta-structure (94%) in mixed dimyristoylphosphatidylcholine/dimyristoylphosphatidic acid (80/20 w/w) small unilamellar vesicles. the fluorescence decay at 344 ... | 1987 | 3318926 |
| [formation and properties of artificial polycistrons containing truncated genes for e. coli tryptophan operon and phage m13 envelope protein]. | using gene fragments encoding the leader peptide of e. coli tryptophane operon (as duplicated fragment hhai-140) or m13 phage coat protein (as taqi-381 or haeiii-1623 fragments) and basing on pds1 family of plasmids, expression vectors have been constructed which contained transcription promoters ptrp, pviii, and pv + pviii, respectively. an artificial gene for human leukocyte interferon alpha 2 (ifn-alpha 2) has been cloned into these plasmids, so that its transcription was a part of polycistro ... | 1987 | 3322290 |
| [design of a hybrid gene coding for the leader sequence of bacillus amyloliquefaciens alpha-amylase and for human proinsulin]. | the chemically synthesized structure gene of human proinsulin was cloned in e. coli on the secretory vector containing regulatory elements of the bacillus amyloliquefaciens alpha-amylase gene. the proinsulin gene was inserted by the ecori site located immediately after the dna area encoding the alpha-amylase signal peptide. the e. coli cells transformed by such a plasmid produced hybrid protein consisting of the alpha-amylase signal peptide, five amino acid residues after the gene mating and hum ... | 1987 | 3326519 |
| characteristics of lactose-fermenting salmonella strains from poland. | in this study 184 lactose-fermenting salmonella strains, collected in the national salmonella centre from the northern and central parts of ponad were examined. epidemiological, serological and biochemical investigations were carried out. apart from this, chemotherapeutic resistance and male-phage sensitivity were determined. most of strains belonged to s. agona serotype (s. typhimurium and s. oranienburg were also presented) which apart from the lactose-fermenting ability retained all the remai ... | 1987 | 3333475 |
| rabbit liver factor d, a poly(thymidine) template stimulatory protein of dna polymerases: purification and characterization. | factor d, a dna binding protein that enhances the activities of diverse dna polymerases with a common restricted set of templates, was initially characterized in mouse liver but has resisted extensive purification. in this paper, we report that a similar stimulatory activity can be obtained in highly purified form from nuclei of rabbit hepatocytes. the rabbit liver protein increases the rates at which several dna polymerases copy sparsely primed natural dna templates and primed synthetic poly(dt ... | 1988 | 3401461 |
| calf thymus dna polymerases alpha and delta are capable of highly processive dna synthesis. | we have demonstrated that calf thymus dna polymerases alpha and delta are capable of highly processive dna synthesis. processivity values between 300 and 2000 nucleotides were observed when poly(da)-oligo(dt) or singly primed single-stranded circular bacteriophage m13 dna at ph 6.0 and 1 mm magnesium chloride was used. these conditions do not correlate with conditions, ph 7.0 and 5 mm magnesium chloride, that support the maximum synthetic rate. lowering the ph and magnesium concentration lowers ... | 1988 | 3401462 |
| oligonucleotide-directed construction of mutations: a gapped duplex dna procedure without enzymatic reactions in vitro. | the gapped duplex dna approach to oligonucleotide-directed construction of mutations (kramer et al. 1984, nucl. acids res. 12, 9441-9456) has been developed further. a procedure is described that makes in vitro dna polymerase/dna ligase reactions dispensable. direct transfection of host bacteria with gddna molecules of recombinant phage m13 plus mutagenic oligonucleotide results in marker yields in excess of 50% (gap size 1640 nucleotides). an important feature incorporated into the mutagenic ol ... | 1988 | 3405755 |
| 5-azido-2'-deoxyuridine 5'-triphosphate: a photoaffinity-labeling reagent and tool for the enzymatic synthesis of photoactive dna. | we have synthesized the photoactive deoxyuridine nucleotide 5-azido-2'-deoxyuridine 5'-triphosphate (5-n3dutp) and used it to synthesize light-sensitive dna by enzymatic incorporation. in the absence of ultraviolet light, 5-n3dutp is a substrate for escherichia coli dna polymerase i. in in vitro dna synthesis reactions using bacteriophage m13 single-stranded dna as the template and 5-n3dutp in place of dttp, a photoactive complementary strand was synthesized by dna polymerase i. the complementar ... | 1986 | 3461438 |
| human delta-aminolevulinate dehydratase: nucleotide sequence of a full-length cdna clone. | two cdnas encoding human delta-aminolevulinate dehydratase (ala-d; porphobilinogen synthase; ec 4.2.1.24), the second enzyme in the heme biosynthetic pathway, were identified, recloned into bacteriophage m13, and sequenced by primer extension. the first clone with an 827-base-pair (bp) pex-ala-d cdna insert, shown to contain dna sequences that were colinear with four bovine ala-d peptide sequences, was used to screen a pkt218 human liver library. a second clone containing a 1200-bp insert was id ... | 1986 | 3463993 |
| [cloning of dna complementary to mrna for proopiomelanocortin from the bovine, rat and human hypophysis. hormonal regulation of proopiomelanocortin mrna in the rat hypophysis]. | cloning of dna and complementary mrna of bovine, rat and human proopiomelanocortin (pomc) was carried out. a structural analysis of the cloned cdna of pomc was performed. using restriction fragments of bovine, rat and human pomc cloned cdna, probes for molecular hybridization based on one-chain bacteriophage m13 were made. using the dot-hybridization technique with labeled [32p] pomc cdna, the effect of 17 beta-estradiol and adrenalectomy on the pomc mrna level in rat hypophysis was studied. the ... | 1987 | 3474031 |
| isolation of lactoferrin cdna from a human myeloid library and expression of mrna during normal and leukemic myelopoiesis. | lactoferrin is a major constituent of polymorphonuclear leukocyte granules and is present in mature neutrophils but not in blasts or promyelocytes. we have isolated a cdna probe for lactoferrin and used it to study the synthesis of lactoferrin mrna by normal and leukemic granulocyte precursors. the probe phl-41 has been subcloned in phage m13 and characterized by restriction endonuclease analysis and nucleic acid sequencing. phl-41 contains approximately 40% of the coding sequence of the lactofe ... | 1987 | 3477300 |
| different dna-binding modes and cooperativities for bacteriophage m13 gene-5 protein revealed by means of fluorescence depolarisation studies. | the binding of the bacteriophage-m13-encoded gene-5 protein to oligo(deoxythymidylic acid)s and m13 dna was studied by means of tyrosyl fluorescence decay and fluorescence anisotropy measurements. the observed fluorescence decays could be described with two exponentials, characterised by the lifetimes tau 1 = 2.2 ns and tau 2 = 0.8 ns respectively. only the amplitude of the longer-lifetime component is influenced by binding of the protein to dna. this indicates that a part of the tyrosyl residue ... | 1986 | 3486763 |
| tobacco mosaic virus replicase and replicative structures. | the rna-dependent rna polymerase (replicase) mediating the replication of tobacco mosaic virus (tmv) has been investigated in a number of laboratories over a period of 20 years. cell-free enzyme preparations have been prepared which can continue the synthesis of nascent complementary rna, initiated in vivo; however, the enzyme does not require, nor does it respond to, exogenous viral rna as a template. the presence in plants of a virus-stimulated, host-encoded rna-dependent rna polymerase (rdrp) ... | 1987 | 3503886 |
| backbone dynamics of a model membrane protein: 13c nmr spectroscopy of alanine methyl groups in detergent-solubilized m13 coat protein. | the filamentous coliphage m13 possesses multiple copies of a 50-residue coat protein which is inserted into the inner membrane of escherichia coli during infection. 13c nuclear magnetic resonance (nmr) spectroscopy has been used to probe the structure and dynamics of m13 coat protein solubilized in detergent micelles. a comparison of backbone dynamics within the hydrophobic core region and the hydrophilic terminal domains was obtained by biosynthetic incorporation of [3-13c]alanine. alanine is d ... | 1986 | 3513830 |
| biosynthesis of reovirus-specified polypeptides. molecular cdna cloning and nucleotide sequence of the reovirus serotype 1 lang strain s4 mrna which encodes the major capsid surface polypeptide sigma 3. | serotype 1 lang strain s4 mrna, which encodes the major capsid surface polypeptide sigma 3 of reovirions, was cloned as a cdna:mrna heteroduplex in escherichia coli using phage m13. a complete consensus nucleotide sequence for s4 mrna has been determined from cdna clones. the lang strain s4 mrna is 1196 nucleotides in length and possesses an open reading frame with a coding capacity of 365 amino acids, sufficient to account for a sigma 3 polypeptide of 41,212 daltons. comparison of the serotype ... | 1986 | 3518713 |
| selection by genetic transformation of a saccharomyces cerevisiae mutant defective for the nuclear uracil-dna-glycosylase. | a coliphage m13 chimer containing the saccharomyces cerevisiae trp1 gene and ars1 replication origin (mpy2) was grown on an ung- dut- strain of escherichia coli. the resulting single-stranded phage dna had 13% of thymine residues substituted by uracil. this dna failed to transform a delta trp1 yeast strain to prototrophy. however, when a mutagenized yeast stock was transformed with uracil-containing single-stranded mpy2 dna, unstable transformants were obtained. after plasmid segregation, about ... | 1986 | 3519585 |
| structure and expression of the alkb gene of escherichia coli related to the repair of alkylated dna. | when the alkb gene of escherichia coli that controls sensitivity of bacteria to methyl methanesulfonate was placed under the control of the lac regulatory region on a multicopy plasmid, the gene product, alkb protein, was overproduced. by monitoring the band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein was purified to near physical homogeneity. an amino-terminal sequence and total amino acid composition of the purified alkb protein were in accord with the amino acid ... | 1986 | 3536913 |
| a sensitive colorimetric detection of virus dna and oncogene. | advantage of cloning probe dna fragment in phage m13 dna was taken to provide a larger single stranded dna as a hybridization probe. high level of direct enzyme labels was introduced via the m13 dna moiety as well as probe dna. a highly sensitive colorimetric detection of virus dna and oncogene was developed. | 1987 | 3548725 |
| screening recombinant clones containing sequences homologous to escherichia coli genes using single-stranded bacteriophage vector. | detection and isolation of escherichia coli clones carrying vectors with foreign dna sequences partially homologous to specific e. coli genes is difficult because denatured dna in the host genome can hybridize with the probe. in this paper we present a procedure which simplifies this task by using bacteriophage m13 as the cloning vector. the procedure takes advantage of the secretory properties of the phage, as well as the property of nitrocellulose membrane to bind protein and single-stranded d ... | 1986 | 3549464 |
| primary structure of an alpha-tubulin gene of physarum polycephalum. | an alpha-tubulin gene of physarum was isolated as a phage-lambda nm1149 recombinant (designated phage-lambda n alpha tu). phage-lambda n alpha tu contained a 4700 base-pair hindiii nuclear dna fragment of an allele of the altb locus of physarum (one of four unlinked alpha-tubulin gene loci). subfragments of the 4700 base-pair insert of phage-lambda n alpha tu were cloned into phage m13 and the nucleotide sequence was determined by the dideoxy chain termination method. the start point of transcri ... | 1987 | 3586027 |
| biosynthesis of reovirus-specified polypeptides. efficiency of expression of cdnas of the reovirus s1 and s4 genes in transfected animal cells differs at the level of translation. | full-length cdnas of the reovirus serotype 1 lang strain s1 and s4 genes were cloned in escherichia coli using bacteriophage m13 and expressed in monkey cos cells under the control of the sv40 late promoter using the eukaryotic expression vector pjc119. the s1-encoded sigma 1 and s4-encoded sigma 3 gene products were expressed in transfected cos cells and were indistinguishable from the authentic sigma 1 and sigma 3 polypeptides synthesized in reovirion-infected cos cells. the relative translati ... | 1987 | 3617502 |
| biosynthesis of reovirus-specified polypeptides. molecular cdna cloning and nucleotide sequence of the reovirus serotype 1 lang strain s2 mrna which encodes the virion core polypeptide sigma 2. | human reovirus serotype 1 lang strain s2 mrna, which encodes the virion inner capsid core polypeptide sigma 2, was cloned as a cdna:mrna heteroduplex in escherichia coli using phage m13. a complete consensus nucleotide sequence was determined. the lang strain s2 mrna is 1331 nucleotides in length and possesses an open reading frame with a coding capacity of 335 amino acids, sufficient to account for a sigma 2 polypeptide of 37,682 daltons. comparison of the serotype 1 lang s2 sequence derived fr ... | 1987 | 3663211 |
| synthesis of a fixed-length single-stranded dna probe by blocking primer extension in bacteriophage m13. | a simple and efficient technique has been developed for preparing radiolabeled single-stranded (ss) probes of determined length and high specific radioactivity. the human beta-globin gene intervening segment ii (ivsii) fragment (0.9-kb) was inserted between the ecori and bamhi sites of m13mp11 and used as a template for ss probe synthesis. the m13 hybridization probe primer (m13 hpp) was annealed to the recombinant m13mp11-beta ivsii template dna. this m13 hpp was next blocked by the enzymatic a ... | 1986 | 3721200 |
| nucleotide sequence identity of mitochondrial dna from different human tissues. | recombinant dna techniques have been used to search for mitochondrial (mt) nucleotide (nt) sequence differences between human tissues within an individual. mtdna isolated from brain, heart, liver, kidney, and skeletal muscle of two different individuals was cleaved with saci and xbai, and then cloned in bacteriophage m13. partial nt sequence determination of 121 independently isolated recombinant m13 clones containing either the cytochrome oxidase subunit iii gene or the d-loop region of human m ... | 1986 | 3744049 |