Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
|---|
| [formation of free radicals and dna breaks after pulsed laser irradiation of dna complexes with intercalating dyes]. | destruction of lambda phage dna is studied under nanosecond pulse laser irradiation (lambda = 355 nm) of dna-dye complexes in solution at 77k (dye--acridine orange, 8-methoxypsoralen, ethidium bromide). free radicals induced by laser radiation are found to participate in dna sugar-phosphate chain scission. it was observed that the quantity of dna double-strand breaks correlated with that of the free radicals and that of oxygen influenced dna laser destruction. | 1990 | 2161260 |
| a herpes simplex virus type 1 latency-associated transcript mutant reactivates with normal kinetics from latent infection. | the herpes simplex virus type 1 (hsv-1) latency-associated transcripts (lats) accumulate in neuronal nuclei of latently infected ganglia. explant reactivation kinetics of lat deletion mutants in the mouse eye model have suggested a role for the lats in the reactivation process. this report describes the construction and characterization of an hsv-1 strain hfem mutant, tb1, disrupted within both copies of the lat gene. tb1 contains a 440-base-pair segment of bacteriophage lambda dna in place of a ... | 1990 | 2161947 |
| single-step labeling of dna using restriction endonucleases and t4 polynucleotide kinase. | dna restriction fragments can be end-labeled in a combined digestion and labeling reaction with restriction endonuclease plus t4 polynucleotide kinase, omitting a dephosphorylation step. both pbr322 and phage lambda dna digests are efficiently labeled in one step, one tube and one incubation. | 1990 | 2162682 |
| cloning and characterization of molecular isoforms of the catalytic subunit of calcineurin using nonisotopic methods. | the cloning and characterization of cdnas for the catalytic subunit of calcineurin (cn) from murine and human brain libraries were carried out using nonisotopic methods. a murine cdna clone encoding a protein of 521 amino acids (mr approximately 58,650) was isolated; overlapping clones established a 3'-untranslated region of 554 base pairs preceding the poly(a) tail. homologous cdnas from human brain showed greater than 92% nucleotide sequence identity in both coding and non-coding regions with ... | 1990 | 2162844 |
| superfamily of uvra-related ntp-binding proteins. implications for rational classification of recombination/repair systems. | a superfamily of proteins encoded by bacterial, phage and eukaryotic genomes and performing a wide range of ntp-dependent functions was delineated by amino acid sequence comparison. the new superfamily brought together bacterial proteins uvra, recf, recn, muth and hexa, t4 phage gp46, t5 phage d13 protein, lambda phage ea59 protein and yeast rad50 protein, all involved in recombination, repair and, in some cases, also in replication of respective genomes, and a family of bacterial and eukaryotic ... | 1990 | 2162963 |
| transposition in shigella dysenteriae: isolation and analysis of is911, a new member of the is3 group of insertion sequences. | twenty-nine clear-plaque mutants of bacteriophage lambda were isolated from a shigella dysenteriae lysogen. three were associated with insertions in the ci gene: two were due to insertion of is600, and the third resulted from insertion of a new element, is911. is911 is 1,250 base pairs (bp) long, carries 27-bp imperfect terminal inverted repeats, and generates 3-bp duplications of the target dna on insertion. it was found in various copy numbers in all four species of shigella tested and in esch ... | 1990 | 2163395 |
| genetic control of the uv-induced sos mutator effect in single- and double-stranded dna phages. | the sos hypothesis postulated that the mutator effect on undamaged dna that generates phage-untargeted mutagenesis (utm) results directly from the mechanism of targeted mutagenesis. reca protein, which stimulates the cleavage of both the lexa repressor and umud protein, and the umudc gene products are required for uv-induced targeted mutagenesis. the use of phage lambda for analyzing uv-induced mutagenesis has permitted a distinction to be made between the mechanisms of targeted and untargeted m ... | 1990 | 2165215 |
| host virus interactions in the initiation of bacteriophage lambda dna replication. recruitment of escherichia coli dnab helicase by lambda p replication protein. | the bacteriophage lambda p protein promoters replication of the phage chromosome by recruiting a key component of the cellular replication machinery to the viral origin. specifically, p protein delivers one or more molecules of escherichia coli dnab helicase to a nucleoprotein structure formed by the lambda o initiator at the lambda replication origin. using purified proteins, we have examined the features of the pivotal host virus interaction between p and dnab. these two proteins interact in v ... | 1990 | 2165499 |
| achilles' heel cleavage: creation of rare restriction sites in lambda phage genomes and evaluation of additional operators, repressors and restriction/modification systems. | a novel technique for the creation of rare restriction sites was described by koob et al. [science 241 (1988) 1084-1086]. this technique, achilles' heel cleavage (ac), relies on the use of a bound repressor molecule to protect only one of many identical restriction sites from a modification methyltransferase that inactivates all other restriction sites. the technique was applied to a small plasmid and shown to work efficiently with two repressor/operator systems: lac repressor/laco operator and ... | 1990 | 2165969 |
| vectors for constructing kan gene fusions: direct selection of mutations affecting is10 gene expression. | we describe several vectors for constructing translational fusions to the kan gene of tn5. fusions are constructed in vitro using multi-copy vectors containing unique cloning sites situated between upstream transcriptional terminators and a downstream kan gene lacking transcriptional and translational start signals. multi-copy fusions can be converted to single-copy chromosomal fusions by in vivo recombination with specific phage lambda vectors and vice versa. we find that kan fusions are often ... | 1990 | 2165970 |
| a grpe mutant of escherichia coli is more resistant to heat than the wild-type. | the grpe gene of escherichia coli is essential for bacteriophage lambda dna replication and is also necessary for host rna and dna synthesis at high temperature. a grpe mutant of e. coli was found to be substantially more resistant to 50 degrees c heat treatment than the wild-type. upon receiving a 42 degrees c heat shock for 15 min, both the wild-type and the grpe mutant became more resistant to heat (i.e. they became thermotolerant). a grpe+ revertant behaved similarly to the wild-type in that ... | 1990 | 2166131 |
| bending and supercoiling of dna at the attachment site of bacteriophage lambda. | integration of the dna of bacteriophage lambda into the chromosome of e. coli depends on the formation of a complex nucleoprotein array at a specific locus on the phage genome, the attachment site. recent work shows how bending of this dna (induced by a specific dna-binding protein), and strain in this dna (induced by supercoiling) contribute to the formation of the nucleoprotein structure. further, there are new insights into the way this structure directs critical events during recombination. | 1990 | 2166364 |
| cloning of three human tyrosine phosphatases reveals a multigene family of receptor-linked protein-tyrosine-phosphatases expressed in brain. | a human brainstem cdna library in bacteriophage lambda gt11 was screened under conditions of reduced hybridization stringency with a leukocyte common antigen (lca) probe that spanned both conserved cytoplasmic domains. cdna encoding a receptor-linked protein-tyrosine-phosphatase (protein-tyrosine-phosphate phosphohydrolase, ec 3.1.3.48), rptpase alpha, has been cloned and sequenced. human rptpase alpha consists of 802 amino acids. the extracellular domain of 150 residues includes a hydrophobic s ... | 1990 | 2169617 |
| molecular structures of two human dna topoisomerase i retrosequences. | we have isolated recombinant lambda-phage clones that contain sequences complementary to the 3' half of the cdna encoding human topoisomerase i (htop1). these lambda clones belong to three distinct classes: class-i clones contain sequences from the active gene located on human chromosome 20. class-ii and class-iii clones contain sequences corresponding to the cdna encoding htop1 from nucleotide (nt) 2208 to 3434 and from nt 1639 to 3434, respectively. these sequences exhibit the characteristic f ... | 1990 | 2170234 |
| dna cleavage during ethanol metabolism: role of superoxide radicals and catalytic iron. | the generation of superoxide and related free radicals and the mobilization of catalytic iron due to ethanol metabolism have been suggested as mechanisms of alcohol-induced liver injury as well as of the increased risk of cancer observed in alcoholics. cleavage of double stranded dna is produced by both free radicals as well as by catalytic iron. the effects of ethanol metabolism on dna cleavage were therefore studied in vitro as well as in vivo in isolated hepatocytes. intactness of double stra ... | 1990 | 2170794 |
| sse8387i, a new type-ii restriction endonuclease that recognizes the octanucleotide sequence 5'-cctgcagg-3'. | a type ii restriction endonuclease designated sse8387i was partially purified from streptomyces sp. 8387. this enzyme cleaved adenovirus 2 dna at three sites, lambda phage dna at five sites, and puc18 and m13mp18 rf dna at one site each, but did not cleave the dnas from pbr322, sv40, or phi x174. sse8387i recognized the octanucleotide sequence 5'-cctgca decreases gg-3', cleaving where shown by the arrow. sse8387i is the first restriction endonuclease to be reported that recognizes an octanucleot ... | 1990 | 2170941 |
| tna transposons can be introduced and maintained in neisseria gonorrhoeae. | in neisseria gonorrhoeae, all penicillinase-specifying plasmids isolated so far share homology with each other and carry a 60% deleted sequence of tna. plasmid phd131, an element isolated from haemophilus ducreyi and carrying an intact copy of the ampicillin resistance transposable element, was introduced from escherichia coli into n. gonorrhoeae by both transformation and conjugative mobilization. plasmids were recovered with no detectable deletion. after their transfer back into e. coli, trans ... | 1990 | 2171110 |
| in vitro transposition of transposon tn3. | transposition of the ampicillin-resistant transposon tn3 was reproduced in vitro using the escherichia coli cell extract. in this cell-free system, we used plasmid dna carrying mini-tn3 as donor and phage lambda dna as target and assayed for ampicillin-resistance transducing phages formed by cointegration of these dna molecules. ampicillin-resistance transducing phages, which were obtained by in vitro packaging of lambda dna after the in vitro transposition reaction, were formed only in the pres ... | 1990 | 2172235 |
| effects of medium quality on the expression of human interleukin-2 at high cell density in fermentor cultures of escherichia coli k-12. | we examined the ability of transformed escherichia coli cells in fermentor cultures to accumulate interleukin-2 (il-2) intracellularly under temperature-regulated control of the phage lambda pl promoter. induction of expression was undertaken at different culture optical densities, and specific il-2 accumulation was found to decrease with increasing cell density at induction. induction at higher culture optical densities was also accompanied by decreased growth during induction and increased ace ... | 1990 | 2180368 |
| identification and characterization of a new escherichia coli gene that is a dosage-dependent suppressor of a dnak deletion mutation. | we report the isolation and characterization of a previously unidentified escherichia coli gene that suppresses the temperature-sensitive growth and filamentation of a dnak deletion mutant strain. introduction of a multicopy plasmid carrying this wild-type gene into a dnak deletion mutant strain rescued the temperature-sensitive growth of the dnak deletion mutant strain at 40.5 degrees c and the filamentation, fully at 37 degrees c and partially at 40.5 degrees c. however, the inability of dnak ... | 1990 | 2180916 |
| genomic organization and physical mapping of the transfer rna genes in escherichia coli k12. | by using a set of 476 ordered dna clones (in lambda phage vector) that covers the entire chromosome of escherichia coli k12, we have made an exhaustive survey of trna genes in the e. coli genome. ultraviolet-irradiated bacteria were separately infected with each of the 476 clones and the rna molecules produced upon infection were labeled with 32p. the labeled trnas were separated by gel electrophoresis and then characterized by fingerprinting analysis. fifty-nine of the 476 clones produced trnas ... | 1990 | 2184240 |
| expression of agrobacterium rhizogenes rolb gene fusions in escherichia coli: production of antibodies against the rolb protein. | expression of the rolb gene of agrobacterium rhizogenes tl-dna is sufficient to trigger root differentiation in transformed plant cells. to investigate the role of rolb in differentiation, a large portion of rolb, comprising about 90% of its c-terminal coding sequence, was cloned into vectors pex34 and pea305 in frame with the truncated n termini of the pl-ms2 phage dna polymerase and, respectively, the ptac-c its phage lambda repressor gene. hybrid proteins were expressed from both fusions and ... | 1990 | 2185136 |
| compilation of dna sequences of escherichia coli (update 1990). | we have compiled the dna sequence data for e.coli available from the genbank and embl data libraries and over a period of several years independently from the literature. this is the second listing replacing and increasing the former listing roughly by one third. after deletion of all detected overlaps a total of 1 248 696 individual bp is found to be determined till the beginning of 1990. this corresponds to a total of 26.46% of the entire e. coli chromosome consisting of about 4,720 kbp. this ... | 1990 | 2185457 |
| expression of the e. coli nadb gene and characterization of the gene product l-aspartate oxidase. | quinolinic acid is synthesized in e. coli by the enzymes l-aspartate oxidase and quinolinate synthase a, the genes of which are named nadb and nada. in our previous work we cloned and characterized the two genes (flachmann, r., kunz, n., seifert, j., gütlich, m., wientjes, f.j., läufer, a. & gassen, h.g. (1988) eur. j. biochem. 175, 221-228). here we report on the expression of the nadb gene under control of the inducible left promoter of the bacteriophage lambda. the yield of the active gene pr ... | 1990 | 2187483 |
| tgatg vector: a new expression system for cloned foreign genes in escherichia coli cells. | a tgatg vector system was developed that allows for the construction of hybrid operons with partially overlapping genes, employing the effects of translational coupling to optimize expression of cloned cistrons in escherichia coli. in this vector system (plasmid ppr-tgatg-1), the coding region of a foreign gene is attached to the atg codon situated on the vector, to form the hybrid operon transcribed from the phage lambda pr promoter. the cloned gene is the distal cistron of this hybrid operon ( ... | 1990 | 2187746 |
| escherichia coli metr mutants that produce a metr activator protein with an altered homocysteine response. | using an escherichia coli lac deletion strain lysogenized with a lambda phage carrying a meth-lacz gene fusion, we isolated trans-acting mutations that result in simultaneous 4- to 6-fold-elevated meth-lacz expression, 5- to 22-fold-lowered mete-lacz expression, and 9- to 20-fold-elevated metr-lacz expression. the altered regulation of these genes occurs in the presence of high intracellular levels of homocysteine, a methionine pathway intermediate which normally inhibits meth and metr expressio ... | 1990 | 2188942 |
| [cloning and regulation of gene expression of ecorv restriction- modification system]. | a number of recombinant plasmids, containing ecorv restriction-modification genes have been constructed. individual genes of this system were introduced into plasmids of various incompatibility groups. promoter regions of genes encoding methylase and restrictase have been cloned and studied. with the use of specialized vector pve8 it was shown that the efficiency of the endonuclease gene promoter is comparable with early lambda phage promoters and produced about 70% of pl efficiency. the efficie ... | 1990 | 2194115 |
| the role of carbohydrate in the function of human plasminogen: comparison of the protein obtained from molecular cloning and expression in escherichia coli and cos cells. | a cdna library was constructed in the phage lambda gt11 from human liver mrna enriched for plasminogen mrna by chromatography on sepharose 4b. a full-length cdna clone of human plasminogen was isolated. the 2.7 kb cdna encoded the entire plasminogen molecule, a signal peptide sequence and two start codons with a 5'-untranslated region of about 80 base pairs. in the 3'-non coding region of 280 base pairs a consensus signal aataaa was found at a distance of 46 base pairs upstream of the poly(a) ta ... | 1990 | 2198941 |
| identification and characterization of a target antigen of a monoclonal antibody directed against eimeria tenella merozoites. | monoclonal antibodies (mab) were produced against eimeria tenella merozoites. a single mab, lpmc-61, was selected because of its ability to bind to merozoites by indirect immunofluorescence assay (ifa) and to inhibit in vitro sporozoite development. mab lpmc-61 reacts with an approx. 10-12-kda merozoite polypeptide in reduced sds-page, but with an approx. 80-kda protein in non-reduced sds-page. the monoclonal recognizes similarly sized polypeptides in e. tenella sporozoites, oocysts and schizont ... | 1990 | 2200963 |
| dna sequence analysis of gamma radiation-induced deletions and insertions at the aprt locus of hamster cells. | gamma radiation-induced gene rearrangements at the chinese hamster ovary cell locus coding for the purine salvage enzyme adenine phosphoribosyl transferase (aprt) consist of both simple deletions and more complex alterations that are presumably the result of multiple strand breaks. to characterize these mutations at the dna sequence level, fragments altered by deletion and insertion mutations were obtained by cloning in lambda phage vectors or by using the polymerase chain reaction. the radiatio ... | 1990 | 2206285 |
| transition of the mrna sequence downstream from the initiation codon into a single-stranded conformation is strongly promoted by binding of the initiator trna. | using an rna footprinting technique, accessible sites on the mrna initiation region bound to the ribosome have been determined. chemical probing experiments have been done both in the presence and absence of the initiator trna with dimethyl sulfate, kethoxal and carbodiimide as reagent probes. as an mrna, a mini-mrna containing the initiation region of bacteriophage lambda gene cro has been used. this region is characterized by a long single-stranded shine-dalgarno (sd) sequence followed by two ... | 1990 | 2207137 |
| construction and characterization of a recombinant murine monoclonal antibody directed against human fibrin fragment-d dimer. | cdna libraries in lambda phage were generated from the murine hybridoma secreting mab-15c5, a monoclonal antibody directed against fragment-d dimer of crosslinked human fibrin [holvoet et al. (1989) thromb. haemostasis 61, 307-313], and clones encoding fragments of the heavy (gamma 1) and the light (kappa) chain were isolated. the kappa-chain cdna was reconstructed from two overlapping clones encoding 20 amino acids of signal sequence and the 214 amino acids of the mature protein chain. the gamm ... | 1990 | 2209622 |
| chromosome iii of saccharomyces cerevisiae: an ordered clone bank, a detailed restriction map and analysis of transcripts suggest the presence of 160 genes. | using lambda phage vector embl4, we isolated 344 clones containing segments of chromosome iii of saccharomyces cerevisiae, analysed their physical structure with eight restriction enzymes and sorted the data in contiguous groups with computer programmes. furthermore, we performed southern hybridizations between the sorted contiguous clone groups and interrelated them into larger groups. in this way, we constructed an ordered clone bank that covers almost the whole of chromosome iii with a single ... | 1990 | 2220074 |
| cloning, expression and sequence analysis of an endolysin-encoding gene of lactobacillus bulgaricus bacteriophage mv1. | the lysa gene specifying an endolysin of lactobacillus delbrueckii subsp. bulgaricus bacteriophage mv1, was cloned and expressed in escherichia coli. the 4.05-kb restriction fragment containing this gene was analysed by restriction and deletion mapping, and by subcloning. the nucleotide sequence of a 1150-bp fragment coding for an active lysin was determined. the lysa gene consists of 585 bp and codes for a protein of a deduced mr of 21,120, which agrees with the size based on in vivo transcript ... | 1990 | 2227453 |
| cloning and sequencing of a cdna for the hemolymph juvenile hormone binding protein of larval manduca sexta. | a cdna for the hemolymph juvenile hormone binding protein (jhbp) of larval manduca sexta has been cloned and sequenced. the jhbp was purified to homogeneity from fifth instar larval hemolymph using gel filtration chromatography, ion exchange chromatography, and preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. polyclonal rabbit antibodies, generated in response to this protein, were used to identify and isolate jhbp cdnas from a fat body expression library in bacteriophage l ... | 1990 | 2246263 |
| characterization of the 5'-flanking region of the rat protein kinase c gamma gene. | the 5'-flanking region of protein kinase c (pkc) gamma gene was identified from a rat liver genomic library in a bacteriophage lambda charon 4a. a 3.6-kilobase (kb) genomic fragment containing the 5'-flanking region, first exon, and first intron was isolated and sequenced. the transcriptional initiation site, identified by s1 mapping and primer extension, was located 243 base pairs upstream from the translational initiation site. promoter activity of a dna segment spanning the 5'-flanking region ... | 1990 | 2246272 |
| molecular cloning of complementary dnas encoding two cationic peroxidases from cultivated peanut cells. | we have isolated, cloned, and characterized two cdnas corresponding to the mrnas for cationic peroxidases synthesized by cultured peanut cells. the first clone was obtained from a phage lambda gt11 library screened with antibodies directed against the major secreted isozyme. its predicted amino acid sequence, deduced from the 1228-base-pair (bp) cdna, revealed a 22-amino acid signal peptide and a 294-amino acid mature protein (mr, 31,228). the second clone was isolated from a lambda gt10 library ... | 1990 | 2247460 |
| further tests of a recombination model in which chi removes the recd subunit from the recbcd enzyme of escherichia coli. | when one of two infecting lambda phage types in a replication-blocked cross is chi + and dna packaging is divorced from the recbcd-chi interaction, complementary chi-stimulated recombinants are recovered equally in mass lysates only if the chi + parent is in excess in the infecting parental mixture. otherwise, the chi 0 recombinant is recovered in excess. this observation implies that, along with the chi 0 chromosome, two chi + parent chromosomes are involved in the formation of each chi + recom ... | 1990 | 2249753 |
| mouse u14 snrna is encoded in an intron of the mouse cognate hsc70 heat shock gene. | mouse u14 snrna (previously designated mouse 4.5s hybrna) is an evolutionarily conserved eukaryotic low molecular weight rna capable of intermolecular hybridization with both homologous and heterologous 18s rrna (1). a single genomic fragment of mouse dna containing the u14 snrna gene(s) has been isolated from a charon 4a lambda phage mouse genomic library and sequenced. results have surprisingly revealed the presence of three u14 snrna-homologous regions positioned within introns 5, 6, and 8 of ... | 1990 | 2251119 |
| saturation and specificity of the lon protease of escherichia coli. | lon is an atp-dependent protease of escherichia coli. the lon mutation has a pleiotropic phenotype: uv sensitivity, mucoidy, deficiency for lysogenization by bacteriophage lambda and p1, and lower efficiency in the degradation of abnormal proteins. all of these phenotypes are correlated with the loss of protease activity. here we examine the effects of overproduction of one lon substrate, sula, and show that it protects two other substrates from degradation. to better understand this protection, ... | 1990 | 2254276 |
| molecular cloning and sequence analysis of cdna encoding human ferrochelatase. | the cdna encoding human ferrochelatase [ec 4.99.1.1] was isolated from a human placenta cdna library in bacteriophage lambda gt11 by screening with a radiolabeled fragment of mouse ferrochelatase cdna. the cdna had an open reading frame of 1269 base pairs (bp) encoding a protein of 423 amino acid residues (mr. 47,833) with alternative putative polyadenylation signals in the 3' non-coding regions and poly (a) tails. amino acid sequencing showed that the mature protein consists of 369 amino acid r ... | 1990 | 2260980 |
| natural plasmid transformation in a high-frequency-of-transformation marine vibrio strain. | the estuarine bacterium vibrio strain di-9 has been shown to be naturally transformable with both broad host range plasmid multimers and homologous chromosomal dna at average frequencies of 3.5 x 10(-9) and 3.4 x 10(-7) transformants per recipient, respectively. growth of plasmid transformants in nonselective medium resulted in cured strains that transformed 6 to 42, 857 times more frequently than the parental strain, depending on the type of transforming dna. these high-frequency-of-transformat ... | 1990 | 2268155 |
| purification of acetoacetate decarboxylase from clostridium acetobutylicum atcc 824 and cloning of the acetoacetate decarboxylase gene in escherichia coli. | in clostridium acetobutylicum atcc 824, acetoacetate decarboxylase (ec 4.1.1.4) is essential for solvent production, catalyzing the decarboxylation of acetoacetate to acetone. we report here the purification of the enzyme from c. acetobutylicum atcc 824 and the cloning and expression of the gene encoding the acetoacetate decarboxylase enzyme in escherichia coli. a bacteriophage lambda embl3 library of c. acetobutylicum dna was screened by plaque hybridization, using oligodeoxynucleotide probes d ... | 1990 | 2268159 |
| dna binding activity of casein kinase ii. | casein kinase ii, an ubiquitous, oligomeric, messenger-independent protein kinase has previously been shown to concentrate in the nuclear compartment when cells are stimulated to proliferate. the present communication reports that purified mammalian ckii interacts with genomic dna preparations in vitro. this interaction led to an apparent activation of the kinase, most likely explained by prevention of its aggregation and subsequent denaturation. binding of ckii was optimum with double stranded ... | 1990 | 2268349 |
| unexpected loss of genomic dna from agarose gel plugs. | intact chromosomal dnas are routinely prepared by embedding cells in agarose plugs before lysis. the large sizes of the genomic dnas cause their retention while other macromolecules diffuse into and out of the gel matrix during lysis, washing and restriction cleavage incubations. however, in an analysis of agarose-embedded chromosomal dnas cleaved with restriction enzymes, fragments larger than 30 kilobases were found to have eluted from the gel plugs. since loss of fragments from gel plugs may ... | 1990 | 2268419 |
| expression of human liver cytochrome p450 iiia4 in yeast. a functional model for the hepatic enzyme. | cytochrome p-450 (p450) nf, a member of the p450 iiia subfamily, is the major contributor to the oxidation of the calcium-channel blocker nifedipine in human liver microsomes. a cdna clone designated nf25 encoding for human p450 nf was isolated from a bacteriophage lambda gt11 expression library [beaune, p. h., umbenhauer, d. r., bork, r. w., lloyd, r. s. & guengerich, f. p. (1986) proc. natl acad. sci. usa 83, 8064-8068]. we have expressed nf25 cdna in saccharomyces cerevisiae using an expressi ... | 1990 | 2269307 |
| a candida albicans homolog of a human cyclophilin gene encodes a peptidyl-prolyl cis-trans isomerase. | a candida albicans cdna and its genomic counterpart were isolated from lambda phage libraries using a human t-cell cyclophilin (cyp) cdna as a hybridization probe. the clones contain a 486-bp open reading frame predicting a 162-amino acid, approx. 18 kda protein which is similar in size to, and which shares 68 and 81% homology with, human t-cell cyp and cytosolic saccharomyces cerevisiae cyp, respectively. northern blots show the presence of a single mrna species of about 800 bp. however, genomi ... | 1990 | 2269432 |
| random silent mutagenesis in the initial triplets of the coding region: a technique for adapting human glutathione reductase-encoding cdna to expression in escherichia coli. | the introduction of random silent mutations into the 5'-coding region of a human cdna as the basis for successful expression in escherichia coli is demonstrated in four steps. (1) plasmid pub200 containing the prpl promoters of phage lambda was found not to serve as an expression vector for a unchanged human glutathione reductase (hgr)-encoding cdna. (2) when this cdna was expressed in a two-cistron context using high-copy-number plasmids, recombinant protein was detected in low yield (0.03% of ... | 1990 | 2269437 |
| mutant of the glutamine transfer rna gene as uga suppressor in escherichia coli. | a uga suppressor derived from a glutamine trna gene of escherichia coli k12 was isolated and characterized. phages carrying the suppressor su+2uga could be obtained only from a hybrid transducing phage, h80ci857psu+2oc, but not from the original transducing phage lambda ci857psu+2oc. by dna sequence analysis, it was found that the su+2 uga suppressor obtained has two mutations; one is in the anticodon (tta----tca), as expected, and the other (c----t) is at the 7th position from the 3' end of trn ... | 1990 | 2270083 |
| role of the cro repressor carboxy-terminal domain and flexible dimer linkage in operator and nonspecific dna binding. | a series of mutations comprising single and multiple substitutions, deletions, and extensions within the carboxy-terminal domain of the bacteriophage lambda cro repressor have been constructed. these mutations generally affect the affinity of repressor for specific and nonspecific dna. additionally, substitution of the carboxy-terminal alanine with several amino acids capable of hydrogen-bonding interactions leads to improved specific binding affinities. a mutation is also described whereby cyst ... | 1990 | 2271592 |
| nucleotide sequence of the gene for the b subunit of human factor xiii. | factor xiii (mr 320,000) is a blood coagulation factor that stabilizes and strengthens the fibrin clot. it circulates in blood as a tetramer composed of two a subunits (mr 75,000 each) and two b subunits (mr 80,000 each). the b subunit consists of 641 amino acids and includes 10 tandem repeats of 60 amino acids known as gp-i structures, short consensus repeats (scr), or sushi domains. in the present study, the human gene for the b subunit has been isolated from three different genomic libraries ... | 1990 | 2271707 |
| [isolation and mapping of arbitrary single copy dna fragment located on human chromosome 11p11-q11]. | a library of genomic dna has been constructed in embl3 lambda phage, from a chinese hamster/human lymphocytes somatic cell hybrid carrying human chromosome 11 and 20. recombinants containing human genomic dna origin can be isolated from the hybrid cell genomic library by using species-specific probe. 8 single copy fragments have been isolated from 13 recombinants. one of them designated as fd11-1 has been identified on chromosome 11 by hybridized it with hybrid cell clone panel and mapped on chr ... | 1990 | 2288761 |
| detection of protein-dna complex formation by time-resolved fluorescence depolarization of bound ethidium bromide. | we introduce the use of time-resolved fluorescence spectroscopy to probe the interaction between gene regulatory proteins and dna. changes in the decay kinetics of fluorescence polarization anisotropy of ethidium bromide bound to dna segments report changes in hydrodynamic volume and shape which occurs upon complex formation between protein and dna. we have used the decay of fluorescence polarization anisotropy as a spectroscopic handle on the interaction between several site-specific dna-bindin ... | 1990 | 2291477 |
| differential localization of type i and type iii procollagen messenger ribonucleic acids in inflamed periodontal and periapical connective tissues by in situ hybridization. | inflammatory lesions of periodontal and periapical connective tissue were studied by in situ hybridization to detect cells responsible for type i and type iii collagen production. formalin-fixed and paraffin-embedded tissue specimens from patients with oral lesions of various stages of inflammation were hybridized with cdna probes specific for human pro alpha 1(i) and pro alpha 1(iii) collagen mrnas, and with bacteriophage lambda dna as a control probe. this technique permitted us to localize fi ... | 1990 | 2296161 |
| characterization and expression of the complementary dna encoding rat histidine decarboxylase. | histamine is a neurotransmitter in the central nervous system and an important modulator of gastric acid secretion, vasomotor control, inflammation, and allergic reactions. in biological systems the formation of histamine from its precursor histidine is catalyzed by the enzyme l-histidine decarboxylase (hdc; l-histidine carboxy-lyase, ec 4.1.1.22). we have cloned hdc-encoding cdna from a fetal rat liver cdna library (phage lambda gt11) have deduced the amino acid sequence from the nucleotide seq ... | 1990 | 2300558 |
| deduced amino acid sequence of mouse blood-coagulation factor ix. | a mouse fetal liver cdna library was screened with a cdna clone encoding human blood coagulation factor ix protein (hbcfix). a bacteriophage lambda clone was isolated and the nucleotide sequence of a 2710-bp insert was determined. an open reading frame of 459 amino acids (aa) was identified within the sequence that has an 80% sequence similarity with hbcfix. the cdna contains a long 3'-untranslated sequence similar to that of bcfix gene from human and canine sources. however, instead of a sequen ... | 1990 | 2323576 |
| human spermidine synthase: cloning and primary structure. | using a synthetic deoxyoligonucleotide mixture constructed for a tryptic peptide of the bovine enzyme as a probe, cdna coding for the full-length subunit of spermidine synthase was isolated from a human decidual cdna library constructed on phage lambda gt11. after subcloning into the eco ri site of pbr322 and propagation, both strands of the insert were sequenced using a shotgun strategy. starting from the first start codon, which was immediately preceded by a gc-rich region including four overl ... | 1990 | 2344393 |
| molecular cloning and expression of rat placental lactogen-i complementary deoxyribonucleic acid. | a full-length cdna clone for rat placental lactogen i (rpl-i) has been isolated from a phage lambda gt11 library containing cdna synthesized from day 11 rat placental mrna. by northern blot analysis the rpl-i cdna clone hybridizes to a 1.0-kilobase placental mrna and appears as early as day 10 of gestation. maximal expression of this mrna was observed in day 11 and 12 placenta, and faint hybridization of the rpl-i cdna was also detected in day 18 to term placenta. in contrast, the mouse clone hy ... | 1990 | 2373051 |
| structure and sequence determination of the gene encoding human salivary statherin. | human statherin (stt) is a low-mr (43 amino acids) acidic phosphoprotein secreted mainly by salivary glands. it acts as an inhibitor of precipitation of ca.phosphate salts in the oral cavity. dna (12.2 kb) was isolated from human genomic phage lambda libraries as a series of overlapping clones, and the nucleotide sequence of the stt-encoding gene (stt) was determined. the transcribed region spans 6.5 kb and contains six exons and five introns. upstream dna (1.6 kb) was also sequenced and a numbe ... | 1990 | 2373369 |
| cloning and expression of clostridium acetobutylicum atcc 824 acetoacetyl-coenzyme a:acetate/butyrate:coenzyme a-transferase in escherichia coli. | coenzyme a (coa)-transferase (acetoacetyl-coa:acetate/butyrate:coa-transferase [butyrate-acetoacetate coa-transferase] [ec 2.8.3.9]) of clostridium acetobutylicum atcc 824 is an important enzyme in the metabolic shift between the acid-producing and solvent-forming states of this organism. the purification and properties of the enzyme have recently been described (d. p. weisenborn, f. b. rudolph, and e. t. papoutsakis, appl. environ. microbiol. 55:323-329, 1989). the genes encoding the two subuni ... | 1990 | 2383002 |
| cloning and nucleotide sequences of crotamine genes. | a cdna library containing snake toxin genes was constructed in bacteriophage lambda by using mrna isolated from the glands of the south american rattlesnake, crotalus durissus terrificus. the first high-density screening of 400,000 plaques for crotamine-containing genes yielded over 800 positives when a labeled cdna probe with sequence homology to crotamine was used. four of these clones with insert sizes from 270 to 400 base pairs were chosen and their inserts subcloned into pgem-3z and sequenc ... | 1990 | 2389256 |
| use of non-radioactive dna probes for detection of campylobacter jejuni and campylobacter coli in stool specimens. | dna probes specific for c. jejuni (pdt1720 containing a 1475 base pair fragment) and for c. jejuni and c. coli (pdt1719 containing a 1845 base pair fragment) were isolated from a bacteriophage lambda gt11 genomic library of c. jejuni, using antiserum prepared against a 46 kda major outer membrane protein of c. jejuni. the two probe-fragments had different restriction maps and were only moderately related by dna hybridization analysis. a non-radioactive labelling kit which consisted of alkaline p ... | 1990 | 2402249 |
| action of recbcd enzyme on cruciform dna. | we tested the hypothesis that recbcd enzyme of escherichia coli resolves pre-existing holliday recombination intermediates by examining the action of the purified enzyme on an open-ended dna cruciform with limited ability to branch migrate. the enzyme cleaved two strands of the cruciform near its base to produce "recombinant" products, with a marked bias in the direction of cleavage. the two nicks necessary to cleave the cruciform were made separately. cruciforms whose four termini were blocked ... | 1990 | 2405161 |
| initiation of dna replication on single-stranded dna templates catalyzed by purified replication proteins of bacteriophage lambda and escherichia coli. | initiation of bacteriophage lambda dna replication at the chromosomal origin depends on the lambda o and p replication proteins. these two viral initiators, together with an escherichia coli protein fraction, promote the replication in vitro of single-stranded circular dna chromosomes such as that of bacteriophage m13. this nonspecific strand initiation reaction, which we have termed the "lambda single-strand replication reaction," has now been established with eight purified proteins, each of w ... | 1985 | 2408273 |
| rna sequence and secondary structure requirements for rho-dependent transcription termination. | the interaction of e. coli termination factor rho with the nascent rna transcript appears to be a central feature of the rho-dependent transcription termination process. based on in vitro studies of the rho-dependent termination of the transcript initiated at the pr promoter of bacteriophage lambda, and on earlier studies, morgan, bear and von hippel (j. biol. chem. 258, 9565-9574, 1983) proposed a model defining the features of a potential binding site for rho protein on transcripts subject to ... | 1985 | 2409526 |
| surface proteins of mycoplasma hyopneumoniae identified from an escherichia coli expression plasmid library. | a genomic library of mycoplasma hyopneumoniae was constructed by cloning random dna fragments approximately 300 base pairs long in a fusion expression plasmid, pex29, containing the n terminus of the phage ms2 polymerase under the control of the pl promoter of phage lambda. clones that produced fusion proteins carrying surface-specific antigenic determinants were identified by using antiserum raised in a pig by intranasal inoculation of viable mycoplasmas. rabbit antisera produced against gel-pu ... | 1985 | 2410363 |
| cloning and overexpression of moloney murine leukemia virus reverse transcriptase in escherichia coli. | a pbr322-derived expression vector, plasmid pkd1, was constructed containing the strong leftward promoter (pl) of bacteriophage lambda, the ribosome-binding site (rbs) of the cii gene of lambda, and a unique downstream ndei restriction site for construction of an atg initiation codon. the section of the pol gene of moloney murine leukemia virus (m-mlv) that codes for reverse transcriptase (rt) was cloned into the ndei site of this vector generating the plasmid prt103. upon thermal induction, enz ... | 1985 | 2412939 |
| isolation of a cdna clone for a catalytic subunit of torpedo marmorata acetylcholinesterase. | we have constructed a cdna library from torpedo marmorata electric organ poly(a+) rna in the lambda phage expression vector lambda gt11. this library has been screened with polyclonal anti-acetylcholinesterase antibodies. one clone, lambda ache1, produced a fusion protein which was recognized by the antibodies and which prevented the binding of native acetylcholinesterase in an enzymatic immune assay. these results indicate that lambda ache1 contains a cdna insert coding for a part of a catalyti ... | 1985 | 2415396 |
| sequence and expression of a human type ii mesothelial keratin. | using mrna from cultured human mesothelial cells, we constructed bacterial plasmids and lambda phage vectors that contained cdna sequences specific for the keratins expressed in these cells. a cloned cdna encoding keratin k7 (55 kd) was identified by positive hybrid selection. southern blot analysis indicated that this sequence is represented only once in the human genome, and northern blot analysis demonstrated that the gene encoding k7 is expressed in abundance in cultured bronchial and mesoth ... | 1985 | 2415537 |
| molecular analysis of several classes of endogenous feline leukemia virus elements. | five recombinant dna clones of endogenous feline leukemia virus-related dna sequences were isolated by screening a lambda phage genomic library of cat placental dna with a probe specific to the gag-pol region of infectious feline leukemia virus. the clones containing retroviral long terminal repeat-like sequences demonstrated the existence of different size classes of endogenous elements in the cat genome, including those of nearly full length in which the gag region is heterogeneous but all of ... | 1985 | 2415715 |
| e. coli nusa protein binds in vitro to an rna sequence immediately upstream of the boxa signal of bacteriophage lambda. | the nusa protein of escherichia coli is a factor which mediates termination of transcription. in this paper, we demonstrate that the nusa protein can bind in vitro to a specific site on the mrna of bacteriophage lambda. several rnas were synthesized by in vitro transcription of truncated lambda dna templates, and the activity of nusa binding to these rnas was examined by a millipore filter-binding assay. rnas containing the sequence immediately upstream of the boxa site were trapped on the filte ... | 1985 | 2416564 |
| formation of transmembrane channels in liposomes during injection of lambda dna. | bacteriophage lambda binds to unilamellar liposomes bearing its receptor protein, lamb, and the lambda dna can be injected into the internal aqueous space. during this process, transmembrane channels are formed in the liposomes which permit the entry and escape of small molecules, but not proteins. the channels are stable and persist after dna injection. | 1986 | 2416751 |
| screening of lambda library for differentially expressed genes using in vitro transcripts. | we present an improved approach to the screening of eucaryotic libraries for differential gene expression. previous techniques have generated probe via the harvesting of cellular poly(a)+ rna and synthesizing labeled cdna probe using reverse transcriptase. in our approach we prepare labeled rna probe via in vitro transcription. unlike cdna preparation, in vitro transcription (i) directly reflects the ongoing rate of gene expression, and (ii) allows one to assess expression of genes whose transcr ... | 1985 | 2418709 |
| rat vitamin d binding protein. determination of the full-length primary structure from cloned cdna. | vitamin d binding protein (dbp) is an abundant serum glycoprotein secreted by the liver which transports vitamin d sterols, binds to actin, and is found on the surface of b-lymphocytes and subpopulations of t-lymphocytes. in the current study, a cdna to rat dbp mrna was cloned from a bacteriophage lambda gt 11 rat liver expression library. this dbp cdna clone was identified by immunoblotting and its identity was confirmed by immunoprecipitation of a 54-kda protein after hybrid-assisted translati ... | 1986 | 2419332 |
| a recurrent dna sequence at sites of protein interaction. | variation in the observed spin lattice relaxation rate (robs) interpreted as proton exchange dominated in sequences corresponding to part of promoters where rna polymerase initiates mrna synthesis has been observed by both patel et al. and reid and co-workers. a higher robs was also seen in the ta pair of the gtg/cac in the sequence corresponding to the lambda phage cro repressor binding site by kyogoku et al. as we pointed out in the introduction, the one case where a three-dimensional structur ... | 1985 | 2422883 |
| sequence of human dna polymerase beta mrna obtained through cdna cloning. | a cdna library from polya+ rna of a human teratocarcinoma cell line in phage lambda gt11 was screened with a fragment of the rat beta-polymerase cdna, lambda pol beta-10, as probe. five positive phage were identified and plaque purified. the cdna of one positive clone selected for detailed study was 1257 bp. this insert was sequenced and found to contain the coding region for beta-polymerase, as well as 163 bp and 137 bp from the 5' and 3' untranslated regions, respectively. the primary structur ... | 1986 | 2423078 |
| the rna component of the bacillus subtilis rnase p. sequence, activity, and partial secondary structure. | the gene defining the catalytic rna component of rnase p in bacillus subtilis 168 was cloned into bacteriophage lambda and plasmid vectors. the nucleotide sequence of the gene and its surroundings was determined from the cloned dna and by directly sequencing or reverse transcribing the rnase p rna. the b. subtilis rnase p rna sequence (400-401 nucleotides) is remarkably different from that of escherichia coli (377 nucleotides) (reed, r. e., baer, m. f., guerrier-takada, c., donis-keller, h., and ... | 1986 | 2423526 |
| brain-specific expression of map2 detected using a cloned cdna probe. | we describe the isolation of a set of overlapping cdnas encoding mouse microtubule associated protein 2 (map2), using an anti-map antiserum to screen a mouse brain cdna expression library cloned in bacteriophage lambda gt11. the authenticity of these clones was established by the following criteria: (a) three non-identical clones each expressing a map2 immunoreactive fusion protein were independently isolated from the expression library; each of these clones cross-hybridized at the nucleic acid ... | 1986 | 2423532 |
| a cloned cdna encoding map1 detects a single copy gene in mouse and a brain-abundant rna whose level decreases during development. | screening of a bacteriophage lambda gt11 cdna expression library with a polyclonal anti-microtubule associated protein (map) antiserum resulted in the isolation of two non-cross-hybridizing sets of cdna clones. one set was shown to encode map2 (lewis, s. a., a. villasante, p. sherline, and n. j. cowan, 1986, j. cell biol., 102:2098-2105). to determine the specificity of the second set, three non-overlapping fragments cloned from the same mrna molecule via a series of "walking" experiments were s ... | 1986 | 2423533 |
| escherichia coli grpe gene codes for heat shock protein b25.3, essential for both lambda dna replication at all temperatures and host growth at high temperature. | we have identified the grpe gene product as the b25.3 heat shock protein of escherichia coli on the following evidence: (i) a protein similar in size and isoelectric point to b25.3 was induced after infection of uv-irradiated bacteria by lambda grpe+ transducing phage, (ii) mutant phage lambda grpe40, isolated by its inability to propagate on grpe280 bacteria, failed to induce the synthesis of the b25.3 protein, and (iii) lambda grpe+ revertants, derived from phage grpe40 as able to propagate on ... | 1986 | 2424889 |
| mutations affecting antigenic determinants of an outer membrane protein of escherichia coli. | the escherichia coli lamb protein is located in the outer membrane. it is both a component of the maltose and maltodextrin transport system, and the receptor for phages lambda and k10. it is a trimer composed of three identical polypeptide chains, each containing 421 residues. six independent mutants have been isolated, in which the lamb protein is altered in its interaction with one or more monoclonal antibodies specific for regions of the protein that are exposed at the cell surface. some of t ... | 1986 | 2426104 |
| cloning of the chicken progesterone receptor. | monospecific antibodies directed against the chicken progesterone receptor (pr) form b were used to screen a randomly primed phage lambda gt11 cdna expression library prepared from size-fractionated chicken oviduct mrna. two independent immunoreactive clones, lambda cpr1 and lambda cpr2, were isolated. antibodies selected from anti-pr form b antiserum on matrices of lambda cpr1 and lambda cpr2 fusion proteins detected two proteins on electrophoretic immunoblots of crude and purified pr preparati ... | 1986 | 2426697 |
| transcription termination at lambda tr1 is mediated by interaction of rho with specific single-stranded domains near the 3' end of cro mrna. | to determine whether e. coli rho protein mediates termination of transcription by interacting with specific segments of the nascent transcript, dna oligonucleotides were used to sequester segments of phage lambda cro mrna in hybrid helices. formation of hybrids was demonstrated with rnaase h assays. oligonucleotides complementary to either of two distinct, single-stranded sequences near the 3' end inhibited rho action at tr1, while oligonucleotides complementary to the sequence between those seg ... | 1986 | 2428503 |
| molecular cloning, expression, and chromosomal localization of the gene encoding a human myeloid membrane antigen (gp150). | dna from a tertiary mouse cell transformant containing amplified human sequences encoding a human myeloid membrane glycoprotein, gp150, was used to construct a bacteriophage lambda library. a single recombinant phage containing 12 kilobases (kb) of human dna was isolated, and molecular subclones were then used to isolate the complete gp150 gene from a human placental genomic dna library. the intact gp150 gene, assembled from three recombinant phages, proved to be biologically active when transfe ... | 1986 | 2428842 |
| use of the phage lambda pl promoter for high-level expression of human interferons in escherichia coli. | 1986 | 2429151 | |
| a sensitive, enzymatic assay for the detection of closely opposed cyclobutyl pyrimidine dimers induced in human diploid fibroblasts. | a sensitive, enzymatic assay has been developed for the detection of closely opposed cyclobutyl pyrimidine dimers induced in uv-irradiated human diploid fibroblasts. in this assay closely opposed dimers are detected as bifilar enzyme-sensitive sites. single-strand incisions are made at the positions of individual pyrimidine dimers through the action of m. luteus pyrimidine dimer-dna glycosylase. incisions at closely opposed dimers, effectively expressed as double-strand breaks, are quantified fr ... | 1986 | 2429177 |
| cdna and amino acid sequences of the cell adhesion protein receptor recognizing vitronectin reveal a transmembrane domain and homologies with other adhesion protein receptors. | cells adhere to vitronectin substrates through a cell surface receptor that recognizes an arg-gly-asp sequence in vitronectin. the receptor is a glycoprotein composed of a 150-kda alpha and a 115-kda beta subunit. the alpha subunit consists of two disulfide-bonded chains of 125 kda and 25 kda. cdna clones were isolated for the alpha subunit of the vitronectin receptor from a phage lambda gt11 expression library made with rna from a human fibroblast cell line, imr-90. the identity of the clones t ... | 1986 | 2430295 |
| overproduction and high-yield purification of phospholipase a from the outer membrane of escherichia coli k-12. | the cloned gene for the outer-membrane-bound phospholipase a from escherichia coli was placed under control of the strong pl promoter of phage lambda. induction of pl resulted in a 250-fold overexpression up to about 2% total cellular protein. this overproduced enzyme was indistinguishable from the wild-type enzyme. a homogeneous phospholiphase a preparation was obtained in high yield from overproducing bacteria, using the zwitterionic detergent c12-sulfobetaine and anion-exchange chromatography ... | 1986 | 2430805 |
| rat androgen-binding protein: evidence for identical subunits and amino acid sequence homology with human sex hormone-binding globulin. | the cdna for rat androgen-binding protein (abp) was previously isolated from a bacteriophage lambda gt11 rat testis cdna library and its identity was confirmed by epitope selection. hybrid-arrested translation studies have now demonstrated the identity of the isolates. the nucleotide sequence of a near full-length cdna encodes a 403-amino acid precursor (mr = 44,539), which agrees in size with the cell-free translation product (mr = 45,000) of abp mrna. putative sites of n-glycosylation and sign ... | 1987 | 2432609 |
| immunological self, nonself discrimination. | the ability of immunodominant peptides derived from several antigen systems to compete with each other for t cell activation was studied. only peptides restricted by a given transplantation antigen are mutually competitive. there is a correlation between haplotype restriction, ability to bind to the appropriate transplantation antigen, and ability to inhibit activation of other t cells restricted by the same transplantation antigen. an exception was noted in that a peptide derived from an antige ... | 1987 | 2433769 |
| nucleotide sequence of the outb locus of bacillus subtilis and regulation of its expression. | the outb gene is one of the genes involved in the process of spore outgrowth in bacillus subtilis. the gene has been cloned in bacteriophage lambda and subcloned in plasmids. we have determined the sequence of 2,553 base pairs around the outb locus. the locus was found to code for a protein of about 30,000 daltons. analysis of the in vivo transcripts from this region by rnase protection experiments revealed the presence of two start sites for transcription. two potential promoters for these tran ... | 1987 | 2435704 |
| serological properties and processing in escherichia coli k12 of ompv fusion proteins of vibrio cholerae. | fusion proteins comprising the amino-terminal 99 amino acids of the bacteriophage ms2 replicase and various portions of ompv a major outer membrane protein of vibrio cholerae were expressed in escherichia coli k12. these fusions were expressed under the control of the pl promoter of bacteriophage lambda, and expression was controlled using a cits repressor. fusions occurring within the secretory signal sequence of ompv gave rise to the production of mature ompv. the efficiency, however, decrease ... | 1986 | 2436027 |
| identification of the vaccinia virus gene encoding nucleoside triphosphate phosphohydrolase i, a dna-dependent atpase. | vaccinia virus encapsidates a dna-dependent atpase known as nucleoside triphosphate phosphohydrolase i (nph i). a bacteriophage lambda gt11 expression library of poxvirus dna was screened with antibodies specific for nph i. positive clones were used to probe restriction fragments of vaccinia virus genomic dna to locate the nph i gene. the identity of the open reading frame (orf) was confirmed by placing it downstream of a bacteriophage t7 promoter, transcribing the orf in vitro, and translating ... | 1987 | 2437324 |
| a procedure for large-scale isolation of rna-free plasmid and phage dna without the use of rnase. | a preparative procedure for the large-scale isolation of plasmid dna without the use of rnase is described. crude plasmid dna is prepared using a standard boiling method. high-molecular-weight rna is removed by precipitation with licl, and low-molecular-weight rna is removed by sedimentation through high-salt solution. the procedure is inexpensive, rapid, simple, and particularly suitable for processing several large-scale preparations simultaneously. a similar procedure has been developed for p ... | 1987 | 2437820 |
| bacteriophage lambda cloning system for the construction of directional cdna libraries. | we have developed a bacteriophage lambda cloning vector, lambda orf8, that can be used for the construction of cdna libraries. the wild-type lambda genome contains five bamhi, five ecori, and seven hindiii restriction sites that have all been removed from the genome of lambda orf8. sites for these endonucleases are present within the multiple cloning site of lambda orf8. we report a method for preparing cdnas that can be cloned in a single orientation in our phage vector. the method utilizes the ... | 1987 | 2438693 |
| expression of japanese encephalitis virus antigens in escherichia coli. | the expression of japanese encephalitis virus (je) cdna in escherichia coli has been used to study the functional organization of the viral genome. je protein coding sequences were expressed in e. coli by subcloning random fragments of cloned cdna (p.c. mcada, p.w. mason, c.s. schmaljohn, j.m. dalrymple, t.l. mason, and m.j. fournier, 1987, virology 158, 348-360) into the bacteriophage lambda gt11 expression vector. over 120 lambda gt11 recombinants expressing viral protein sequences as beta-gal ... | 1987 | 2438844 |
| a single base change in the shine-dalgarno region of 16s rrna of escherichia coli affects translation of many proteins. | a single base mutation was constructed at position 1538 of escherichia coli 16s rrna, changing a cytidine to a uridine. this position is in the shine-dalgarno region, thought to be involved in base-pairing to mrna during initiation of protein synthesis. the mutation was constructed by using a synthetic oligodeoxynucleotide that differs in sequence by one base from the wild-type sequence of 16s rrna. this oligonucleotide was used as a primer on single-stranded dna of phage m13, into which was clo ... | 1987 | 2440027 |
| isolation of several cdnas encoding yeast peroxisomal enzymes. | several candidate clones carrying partial cdnas for yeast peroxisomal enzymes, such as catalase, carnitine acetyltransferase, isocitrate lyase, malate synthase and acyl-coa oxidase, were efficiently isolated at a single plating from a phage lambda gt11 recombinant cdna library prepared with poly(a)-rich rna from an n-alkane-grown yeast, candida tropicalis, with a mixture of antibodies against the respective purified enzymes. among them, one candidate clone carrying partial cdna for catalase was ... | 1987 | 2440725 |
| bleomycin-induced mutagenesis in repackaged lambda phage: base substitution hotspots at the sequence c-g-c-c. | dna isolated from lambda phage was treated with bleomycin a2 plus fe2+. the bleomycin-damaged dna was added to lambda packaging extracts and the resulting phage were grown in sos-induced e. coli. under these conditions, treatment of the dna with 0.8 microm bleomycin reduced the viability of the repackaged phage to 3% and increased the frequency of clear-plaque mutants in the progeny by a factor of 16. bleomycin-induced mutations which mapped to the dna-binding domain of the ci gene were subjecte ... | 1987 | 2442605 |
| an improved negative staining technique using a thin quartz membrane as sample support. | a negative staining technique is presented based on the use of 40-60 nm quartz membrane supported by a silicon grid. the quartz membrane is fabricated by thermal growth of silicon dioxide on a silicon substrate followed by an anisotropic silicon etching step giving rectangular holes in the silicon substrate. the hydrophilic membrane is shown to be ideally suited for negative staining due to its spreading characteristics, homogeneity, heat resistance and mechanical stability. micrographs of phage ... | 1987 | 2442861 |