Publications

the bacillus subtilis addab genes are fully functional in escherichia coli.	an escherichia coli recbcd deletion mutant was transformed with plasmids containing the bacillus subtilis add genes. the transformants had relatively high atp-dependent exonuclease- and atp-dependent helicase activities, and their viability, the ability to repair u.v.-damaged dna and the recombination in conjugation were nearly completely restored. the b. subtilis add enzyme did not show chi-activity in phage lambda recombination. the individual b. subtilis add proteins were not able to form an ...	1993	8387145
identification and baculovirus expression of the vp4 protein of the human group b rotavirus adrv.	a complete cdna copy of the fourth rna segment of the human group b rotavirus adult diarrheal rotavirus (adrv) has been cloned into lambda phage and excised into plasmid psk bluescript. gene segment 4 contains 2,303 bases and encodes one long open reading frame beginning at base 16 and terminating at base 2263. the encoded protein contains 749 amino acids, with a calculated molecular mass of 84.4 kda and a pi of 6.1. gene 4 cdna was inserted into a recombinant baculovirus via homologous recombin ...	1993	8386274
sequence relationship of retrotransposable elements r1 and r2 within and between divergent insect species.	r1 and r2 are retrotransposable elements that integrate at specific sites in the 28s ribosomal rna (rrna) genes of bombyx mori and drosophila melanogaster. we have previously shown that most insect species contain insertions in their 28s genes at the r1 and/or r2 site. we have sequenced the 3' half of r1 and r2 elements from three additional insect species: the fungus gnat, sciara coprophila (diptera); the japanese beetle, popillia japonica (colleoptera); and the parasitic wasp, nasonia vitripen ...	1993	8383793
antibodies to the human gamma 2 subunit of the gamma-aminobutyric acida/benzodiazepine receptor.	a gamma-aminobutyric acida (gabaa) receptor (gabaar) gamma 2 subunit (short form) was cloned from an adult human cerebral cortex cdna library in bacteriophage lambda gt11. the 261-bp intracellular loop (il) located between m3 and m4 was amplified using the polymerase chain reaction and inserted into the expression vectors lambda gt11 and pgex-3x. both beta-galactosidase (lacz) and glutathione-s-transferase (gst) fusion proteins containing the gamma 2il were purified, and a rabbit antibody to the ...	1993	8382267
overexpression, purification, dna binding, and dimerization of the escherichia coli uvrd gene product (helicase ii).	we have subcloned the escherichia coli uvrd gene under control of the inducible phage lambda pl promoter and report a procedure for the large-scale purification of helicase ii protein. yields of approximately 60 mg of > 99% pure helicase ii protein, free of detectable nuclease activity, are obtained starting from 250 g of induced e. coli cells containing the overexpression plasmid. overproduction of helicase ii protein at these levels is lethal in e. coli. the extinction coefficient of helicase ...	1993	8380701
rna polymerase idling and clearance in gal promoters: use of supercoiled minicircle dna template made in vivo.	we have developed an in vivo system to engender supercoiled "minicircle" dna containing a single promoter by using the integrative recombination system of bacteriophage lambda. the resulting minicircle templates allow quantitative analysis of the stages of transcription initiation from a promoter, including synthesis of both full-length and aborted transcripts in the same reactions under physiological conditions. we have used such minicircle dna templates to study in vitro transcription of the e ...	1993	8380640
dramatic improvements in viral inactivation with brominated psoralens, naphthalenes and anthracenes.	amino or polyamino derivatives of naphthalene (n-h), anthracene (a-h) and 8-alkoxypsoralen (psr-h) were prepared along with their monobrominated analogs (n-br, a-br and psr-br). the ammonium salts of these compounds are all water soluble and bind strongly to calf thymus dna and to lambda phage, a double-helical dna, protein-coated virus. binding of the sensitizer to dna occurs, presumably by a mixture of hydrophobic, intercalative and electrostatic interactions. relative binding constants to cal ...	1993	8378434
the effect of formalin fixation on restriction endonuclease digestion of dna and pcr amplification.	the effect of formalin fixation on dna and on polymerase chain reaction (pcr) amplification was investigated. lambda phage dna fixed in buffered formalin showed incomplete digestion on restriction endonuclease treatment. the resistance to restriction digestion was dependent on the temperature of fixation, but not affected by salt concentration of the fixative. lambda phage dna fixed in unbuffered formalin showed poor pcr amplification due to degradation of dna during fixation. lambda phage dna f ...	1993	8378178
effect of coliphage lambda p gene mutations on the stability of the lambda o protein, the initiator of lambda dna replication.		1993	8372559
mutagenesis of the p-loop motif in the atp binding site of the reca protein from escherichia coli.	using a combinatorial cassette mutagenesis procedure we have introduced a number of mutations into 10 codons that define the p-loop motif within the atp binding site of the escherichia coli reca protein. the recombinational proficiency of the reca mutants was determined using three genetic assays: survival in the presence of 4-nitroquinoline-1-oxide, survival following uv irradiation and the ability to support plaque formation by a red-gam-chi+ lambda phage. while no amino acid substitutions wer ...	1993	8371266
characterization and overexpression of the pem gene encoding pectin methylesterase of erwinia chrysanthemi strain 3937.	the pem gene encoding the pectin methylesterase (pme) of erwinia chrysanthemi strain 3937 was subcloned and its nucleotide sequence determined. the gene contains an open reading frame of 1098 bp and codes for a protein of 366 amino acids (aa). the mature 37-kda form of the protein is 342 aa long and has a calculated isoelectric point of 9.64. a plasmid was constructed to overproduce pme: a dna fragment carrying pem was amplified by the polymerase chain reaction and cloned downstream from the pl ...	1993	8370537
a cryptic promoter in the o(r) region of bacteriophage lambda.	a cryptic promoter, designated p alpha, initiates transcription within the o(r) region of bacteriophage lambda. transcription from p alpha proceeds in the direction of the ci repressor gene from sites 46 and 48 bp preceding the prm transcription start site. p alpha is likely to compete with both pr and prm for formation of open complexes, since it is only active when pr is mutated and can be suppressed by mutations that increase prm activity. in addition, transcription initiation at p alpha is b ...	1993	8366050
an attempt to substitute the cell binding domain of human fibronectin in lambda phage j protein: computer design and expression.	we superimposed hydropathic indexes of the human fibronectin cell binding domain (cbd) on the lambda phage j protein by computer, and substituted 22 amino acids from the fibronectin cbd for a part of the lambda phage j protein. the fibronectin cell binding domain -arg-gly-asp-ser- (-rgds-) localizes at the junction of hydrophobicity and hydrophilicity. we selected a similar hydrophobic-to-hydrophilic junction in the j protein region for substitution. this junction corresponds to 150 bp of the ps ...	1993	8364096
[calorimetric studies of the effect of amino acid replacements 16gln-leu and 26tyr-asp on the structural organization and stability of the cro-repressor from phage lambda].	the scanning calorimetry technique has been used to study the influence of amino acid substitutions on thermodynamic parameters of formation and stabilization of a cooperative structure of cro repressor of bacteriophage lambda molecule. it is shown that substitutions 16gln-leu and 26tyr-asp enhances the protein molecule stability by 32 degrees c as compared with the wild-type protein. it is also demonstrated that the denaturation enthalpy of the mutated cro differs slightly from that of the wild ...	1993	8361487
high affinity, thyroid-specific human autoantibodies displayed on the surface of filamentous phage use v genes similar to other autoantibodies.	autoantibodies to thyroid peroxidase (tpo) are characteristic of thyroid inflammation in autoimmune thyroid disease. we have used the phage display, h and l chain combinatorial cdna library approach to clone, from thyroid-infiltrating b cells, six new human fab autoantibodies with high affinities (approximately 10(-10) m) for tpo. this library, in the pcomb3 vector, was screened with viable, stably transfected chinese hamster ovary cells expressing human tpo on their surface. the h and l chain g ...	1993	8360495
human kappa chain expression in a lambda phage vector: methods of isolating amplified cdna affects cloning efficiency.	three common methods of isolating amplified dna were evaluated for their effects on cloning efficiency in a phage vector designed to express human kappa chain. the "glass milk" technique gave higher cloning efficiency and protein expression than phenol-chloroform extraction or microfiltration. this shows that the quality of amplified cdna should be considered when studying the human antibody repertoire using this vector system.	1993	8357956
targeted mutagenesis of dna using triple helix-forming oligonucleotides linked to psoralen.	oligonucleotides can bind as third strands of dna in a sequence-specific manner in the major groove in homopurine/homopyrimidine stretches in duplex dna. here we use a 10-base triplex-forming oligonucleotide linked to a psoralen derivative at its 5' end to achieve site-specific, targeted mutagenesis in an intact, double-stranded lambda phage genome. site-specific triplex formation delivers the psoralen to the targeted site in the lambda dna, and photoactivation of the psoralen produces adducts a ...	1993	8356097
modulation of p(rm) activity by the lambda pr promoter in both the presence and absence of repressor.	when the transcription startsites of the phage lambda promoters prm and pr are separated by 82 bp (the wild-type spacing), mutating pr increases the rate of open complex formation at prm at all rna polymerase (rnap) concentrations tested in vitro. this is reflected in a fourfold increase in kappa f (the rate constant for isomerization of closed to open complexes) and a threefold decrease in kb (the equilibrium constant for formation of closed complexes). these effects of mutating pr resemble qua ...	1993	8355271
capillary zone electrophoresis of large dna.	capillary zone electrophoresis (cze) of dna 23.1 to 48.5 kb in length in polyacrylamide solutions of several concentrations provides evidence for polymer concentration and dna length-dependent stretching and orientation of these species and suggests an effective separation at a polymer concentration of about 0.6%. applying a 0.1% polyacrylamide concentration to the lambda-phage dna ladder, at least 5 components are separated; separation improves with lowering of the field strength to 2 v/cm and, ...	1993	8354238
tumor-specific overexpression of a novel keratinocyte lipid-binding protein. identification and characterization of a cloned sequence activated during multistage carcinogenesis in mouse skin.	differential screening of cdna libraries from chemically induced malignant mouse skin squamous cell carcinomas (sccs) identified sequences, including one called mal1, that were up-regulated in their expression at both the benign papilloma and the malignant scc stages during tumor development. the mal1 plasmid cdna clone was used to screen lambda phage cdna libraries made from chemically induced papillomas and sccs. two size classes (655 and 933 nucleotides excluding the poly(a) tail) of full-len ...	1993	8349619
characterization of a temperature-sensitive mutant of salmonella typhimurium defective in apolipoprotein n-acyltransferase.	on screening 440 temperature-sensitive (ts) mutants of salmonella typhimurium, a mutant strain se5312 which accumulated apolipoprotein (alp) at 42 degrees c was identified. in vitro assay of apolipoprotein n-acyltransferase activity indicated that the mutant cell envelope contained reduced activity as compared to the wild-type strain. transduction with a mud-p22 mapping set placed the ts mutation to 14-17 min region of the s. typhimurium chromosome. p22 transduction using transposon insertions i ...	1993	8344936
characterization of the schistosoma mansoni gene encoding the glycolytic enzyme, triosephosphate isomerase.	the complete gene encoding schistosoma mansoni triosephosphate isomerase (tpi) was isolated from a lambda phage genomic library on 2 overlapping clones. these genomic clones have been characterized by restriction mapping and dna sequencing of the 5' flanking region, the exons, the intron boundaries and the polyadenylation addition site. s. mansoni tpi is encoded by 6 exons spanning a region of about 12 kb. the 5 introns are located at positions precisely analogous to those of mammalian tpi genes ...	1993	8341322
a switch in translation mediated by an antisense rna.	antisense rnas regulate expression of target genes in a variety of ways--transcription termination, translation initiation, and mrna stability. we describe a case in which the target gene encodes two polypeptides, and antisense rna causes a switch in its translation by selectively inhibiting synthesis of one of the polypeptides. bacteriophage p22 is a temperate salmonella phage; in the prophage state it expresses only a handful of its genes. one of these genes, sieb, aborts the lytic development ...	1993	8339929
superinfection exclusion (sieb) genes of bacteriophages p22 and lambda.	the superinfection exclusion gene (sieb) of salmonella phage p22 was mapped with phage deletion mutants. the dna sequence in the region was reexamined in order to find an open reading frame consistent with the deletion mapping. several discrepancies with the previously published sequence were discovered. the revised sequence revealed a single open reading frame of 242 codons with six likely translation initiation codons. on the basis of deletion and amber mutant phenotypes, the second of these s ...	1993	8335629
the 92-min region of the escherichia coli chromosome: location and cloning of the ubia and alr genes.	a cosmid (pnd320) bearing 42.5 kb of escherichia coli chromosomal dna, including the genes between xyle and ssb near minute 92 on the linkage map, was isolated by selection for complementation of a dnab mutation. known nucleotide (nt) sequences were used to align restriction maps in this region to the physical map of the chromosome (coordinates 4319.5 to 4362 kb), and to locate precisely and define the orientations of 19 genes. predicted physical linkage of sequenced genes across unsequenced gap ...	1993	8335265
sequence of a variant shiga-like toxin type-i operon of escherichia coli o111:h-.	pcr amplification was used to screen faecal isolates of escherichia coli from a 12-month-old boy with haemolytic uraemic syndrome for the presence of shiga-like toxin (slt)-encoding genes. one isolate, belonging to serotype o111:h-, was positive for slt-i by this method. uv induction indicated that the strain was lysogenic for a lambdoid bacteriophage, but this did not encode the toxin. southern hybridization analysis of chromosomal dna revealed that the slt-i gene was located on an 8.5-kb ecori ...	1993	8335264
cloning and sequencing of the haemophilus influenzae aroa gene.	this study was designed to clone the aroa gene (encoding 5-enolpyruvylshikimate-3-phosphate synthase [epsps]) from haemophilus influenzae using a mixed-primer polymerase chain reaction (pcr). the mixed primers were based on a back translation of previously reported blocks of amino acid homology between different cloned epsps proteins. the pcr-produced probe from h. influenzae dna was used to identify a recombinant lambda phage from which the rest of h. influenzae aroa was subcloned and sequenced ...	1993	8335255
expression of the rz gene and the overlapping rz1 reading frame present at the right end of the bacteriophage lambda genome.	the rz lysis gene of bacteriophage lambda was cloned into the expression vectors, pt7-3 and pt7-7. the recombinant plasmids expressed either a protein of an unexpected 6.5-kda size (pt7-3h and psb54) or two proteins of 6.5 and 17.2 kda (pbh21). the 6.5-kda protein alone did not complement the lysis defect of the lambda rz mutant; hence, this protein was not the rz gene product. complementation observed as a result of pbh21 expression thus can be ascribed to the 17.2-kda protein, which agrees wit ...	1993	8335247
a pcr-based method for high stringency screening of dna libraries.	a rapid method for cloning genomic dna utilizing a pcr-based screening protocol is described. a murine genomic library in lambda phage was subdivided into 64 wells, each containing 1000 clones, and propagated in bacteria. amplified phage from each of 8 wells across columns, and each of 8 wells down rows, were pooled. the pooled phage were screened for the presence of murine m-csf dna by pcr using specific oligonucleotide primers. a single well that contained an m-csf genomic clone was identified ...	1993	8332459
transcriptional antitermination.	antiterminator proteins control gene expression by recognizing control signals near the promoter and preventing transcriptional termination which would otherwise occur at sites that may be a long way downstream. the n protein of bacteriophage lambda recognizes a sequence in the nascent rna, and modifies rna polymerase by catalysing the formation of a stable ribonucleoprotein complex on its surface, whereas the lambda q protein recognizes a sequence in the dna. these mechanisms of antitermination ...	1993	8332211
induction of hepatic mutations in laci transgenic mice.	transgenic b6c3f1 and c57bl/6 mice containing a lambda shuttle vector that carries a laci target and an alpha lacz reporter gene have been constructed for use in in vivo mutagenesis assays. after chemical treatment of mice carrying the laci target gene, genomic dna is isolated and the shuttle vector is recovered by exposing the dna to lambda phage packaging extracts in vitro. mutations in the laci target gene that inactivate the repressor gene allow expression of the alpha lacz reporter gene, re ...	1993	8332090
complete structure of the human gc gene: differences and similarities between members of the albumin gene family.	the sequence of the human gc gene, including 4228 base pairs of the 5'-flanking region and 8514 base pairs of the 3' flanking region (55,136 in total), was determined from five overlapping lambda phage clones. the sequence spans 42,394 base pairs from the cap site to the polyadenylation site, and it reveals that the gene is composed of 13 exons, which are symmetrically placed within the three domains of the gc protein. the first exon is partially untranslated, as is exon 12, which contains the t ...	1993	8325650
structure of the gene encoding mouse adipose differentiation-related protein (adrp).	adipose differentiation-related protein (adrp) is a novel 50-kda membrane-associated protein whose message levels are induced rapidly and maximally after triggering adipocyte differentiation. the gene encoding mouse adrp has been isolated and characterized from four overlapping lambda phage clones. the gene spans 14 kb and contains 8 exons and 7 introns. exons range in size from 50 to 696 bp and intron sizes range from 87 bp to 4.3 kb. major and minor transcription initiation sites were determin ...	1993	8325636
high-level expression, purification, and enzymatic characterization of full-length thermus aquaticus dna polymerase and a truncated form deficient in 5' to 3' exonuclease activity.	the thermus aquaticus dna polymerase i (taq pol i) gene was cloned into a plasmid expression vector that utilizes the strong bacteriophage lambda pl promoter. a truncated form of taq pol i was also constructed. the two constructs made it possible to compare the full-length 832-amino-acid taq pol i and a deletion derivative encoding a 544-amino-acid translation product, the stoffel fragment. upon heat induction, the 832-amino-acid construct produced 1-2% of total protein as taq pol i. the induced ...	1993	8324500
structure of the nifq gene from enterobacter agglomerans 333 and its overexpression in escherichia coli.	the nifq gene, involved in early stages of iron-molybdenum cofactor (femo-co) biosynthesis, was identified downstream of the nifb and niff genes of enterobacter agglomerans. this gene was cloned and its nucleotide sequence determined. the amino acid sequence, as deduced from the nucleotide sequence, revealed an accumulation of cysteine amino acid residues at the c-terminal end of the protein. the cysteine cluster showed the following consensus sequence cys-x4-cys-x2-cys-x5-cys, which is a typica ...	1993	8316214
viral protein kinases and protein phosphatases.	certain large dna viruses (e.g. herpesviruses and poxviruses) encode proteins related to cellular protein-serine/threonine kinases, and hepatitis b virus and vesicular stomatitis virus may encode structurally different protein kinases. other viruses activate cellular protein kinases, e.g. interferon-induced eukaryotic initiation factor-2 kinase, growth factor-induced kinases and protein kinases that regulate mitosis. protein phosphatases are encoded by vaccinia virus and bacteriophage lambda and ...	1993	8309996
an analogue of the dnaj molecular chaperone in escherichia coli.	escherichia coli dnaj functions as a typical molecular chaperone in coordination with other heat shock proteins such as dnak and grpe in a variety of cellular processes. in this study, it was found that e. coli possesses an analogue of dnaj, as judged from not only its primary structure but also its possible function. this protein, named cbpa (for curved dna-binding protein), was first identified as a dna-binding protein that preferentially recognizes a curved dna sequence. cloning and nucleotid ...	1994	8302830
nuclear assembly with lambda dna in fractionated xenopus egg extracts: an unexpected role for glycogen in formation of a higher order chromatin intermediate.	crude extracts of xenopus eggs are capable of nuclear assembly around chromatin templates or even around protein-free, naked dna templates. here the requirements for nuclear assembly around a naked dna template were investigated. extracts were separated by ultracentrifugation into cytosol, membrane, and gelatinous pellet fractions. it was found that, in addition to the cytosolic and membrane fractions, a component of the gelatinous pellet fraction was required for the assembly of functional nucl ...	1994	8294509
overproduction of the toxic protein, bovine pancreatic dnasei, in escherichia coli using a tightly controlled t7-promoter-based vector.	a synthetic gene coding for bovine pancreatic dnasei has been cloned under the control of a t7 promoter present on the plasmid pet11. this construct yields a stable escherichia coli transformant only when transcription from this promoter is tightly controlled. production of recombinant dnasei (rednasei) is achieved by infection of the cells with a mutant lambda phage, ce6, which carries the gene encoding t7 rna polymerase. induced bacterial cultures yield in excess of 2 mg per litre of rednasei ...	1993	8294027
molecular genetics of cryptopleurine resistance in saccharomyces cerevisiae: expression of a ribosomal protein gene family.	the saccharomyces cerevisiae cry1 gene encodes the 40s ribosomal subunit protein rp59 and confers sensitivity to the protein synthesis inhibitor cryptopleurine. a yeast strain containing the cry1-delta 1::ura3 null allele is viable, cryptopleurine sensitive (crys), and expresses rp59 mrna, suggesting that there is a second functional cry gene. the cry2 gene has been isolated from a yeast genomic library cloned in bacteriophage lambda, using a cry1 dna probe. the dna sequence of the cry2 gene con ...	1993	8293976
structure of the channel catfish (ictalurus punctatus) growth hormone gene and its evolutionary implications.	a dna fragment of 1.6 kilo base pairs (kb), encoding part of the channel catfish (ictalurus punctatus) growth hormone (gh) gene, was generated by the polymerase chain reaction (pcr) using 2 degenerate synthetic oligonucleotides (30 and 33 mer) derived from the n- and c-terminal amino acid sequences of the catfish gh polypeptide as amplification primers and with catfish genomic dna as a template. this dna fragment was used as a probe for the isolation of a catfish gh gene from a genomic library c ...	1993	8293072
cloning of cdnas for m-phase phosphoproteins recognized by the mpm2 monoclonal antibody and determination of the phosphorylated epitope.	the mpm2 monoclonal antibody binds to a phospho amino acid-containing epitope present on more than 40 proteins of m-phase eukaryotic cells. we have developed a technique for cloning cdnas encoding mpm2-reactive phosphoproteins from bacteriophage lambda expression libraries. proteins from phage plaques were absorbed to nitrocellulose filters, phosphorylated by m-phase kinases, and screened for mpm2 binding. partial-length cdnas encoding two mpm2-reactive proteins termed mpm2-reactive phosphoprote ...	1994	8290587
mapping the functional domains of bacteriophage lambda integrase protein.	bacteriophage lambda encodes a site-specific recombination system that promotes the movement of the phage genome into and out of the host bacterial chromosome. the phage-encoded integrase (int) is composed of 356 amino acid residues and carries out the required strand exchanges by means of a type i topoisomerase activity. int also contains two distinct dna-binding domains that interact with two different, specific sequences (arm-type and core-type sites) on dna. in order to help understand the m ...	1994	8289327
cloning in a bacteriophage lambda vector for the display of binding proteins on filamentous phage.	we have combined the efficiency and ease of use of bacteriophage lambda vectors with the power of phage display screening technology to create surfzap. the use of bacteriophage lambda allows the construction of large lambda expression libraries, which are rapidly and efficiently converted to stable plasmid libraries by mass excision. in surfzap, clones are expressed as fusions with amino acids 198-406 of the m13 minor coat protein (cpiii) and are displayed on the surface of filamentous phage. wh ...	1993	8282204
structure and polymorphisms of the human annexin iii (anx3) gene.	a 75-kb region encompassing the human annexin iii (anx3) gene on chromosome 4q21 was characterized from directly amplified genomic dna and from six genomic clones in phage lambda (lambda anx3-1 to lambda anx3-6). the gene was mapped with restriction enzymes bamhi, ecori, hindiii, saci, and xbai, and primers were developed for nine sequence-tagged sites throughout the gene. the transcribed region spans 58 kb and contains 12 introns ranging from 0.3 to 19.1 kb and 13 exons ranging from 53 to 374 b ...	1993	8276419
from the double-helix to novel approaches to the sequencing of large genomes.	elucidation of the structure of dna by watson and crick [nature 171 (1953) 737-738] has led to many crucial molecular experiments, including studies on dna replication, transcription, physical mapping, and most recently to serious attempts directed toward the sequencing of large genomes [watson, science 248 (1990) 44-49]. i am totally convinced of the great importance of the human genome project, and toward achieving this goal i strongly favor 'top-down' approaches consisting of the physical map ...	1993	8276270
target of the transcriptional activation function of phage lambda ci protein.	activation of transcription initiation by the ci protein of phage lambda is thought to be mediated by a direct interaction between cl and rna polymerase at the prm promoter. two negatively charged amino acid residues in the dna binding domain of ci play a key role in activation, suggesting that these residues contact rna polymerase. the subunit of rna polymerase involved was identified by selecting polymerase mutants that restored the activation function of a mutant form of ci protein. although ...	1994	8272867
clusters of interspersed repeated dna sequences in the rice genome (oryza).	we have characterized a repeated dna sequence (rtl122) from rice (oryza sativa l.) with respect to its organization in the rice genome and its distribution among rice and other plants. the results indicate that the rtl122 sequence is interspersed in the rice genome and limited to the genus oryza. it is highly polymorphic and can be used to fingerprint rice varieties. a structure was observed in which several repeated sequences were clustered in dna regions of 15-20 kb. we characterized three bac ...	1993	8270205
swift identification of recombinant proteins from lambda-phage libraries without resorting to liquid cultures or lysogens.		1993	8267966
isolation and characterization of a saccharomyces cerevisiae peptide transport gene.	we have cloned and characterized a saccharomyces cerevisiae peptide transport gene (ptr2) isolated from a genomic dna library by directly selecting for functional complementation of a peptide transport-deficient mutant. deletion and frameshift mutageneses were used to localize the complementing activity to a 3.1-kbp region on the transforming plasmid. dna sequencing of the complementing region identified an open reading frame spanning 1,803 bp. the deduced amino acid sequence predicts a hydropho ...	1994	8264579
phage lambda beta protein, a component of general recombination, is associated with host ribosomal s1 protein.	we have purified phage lambda beta protein produced by a recombinant plasmid carrying bet gene and confirm that it forms a complex with a protein of relative molecular mass 70 kda. therefore, beta protein, a component of general genetic recombination, is associated with two functionally diverse complexes; one containing exonuclease and the other 70 kda protein. using a number of independent methods, we show that 70 kda protein is the ribosomal s1 protein of e. coli. further, the association of 7 ...	1993	8260932
an opsin homologue in the retina and pigment epithelium.	the aim of this project was to investigate the retinal pigment epithelium (rpe) at the molecular level by identification of novel rpe-specific cdnas that may encode proteins of signal transduction pathways or other proteins that are expressed preferentially in the rpe.	1993	8258527
the role of cosb, the binding site for terminase, the dna packaging enzyme of bacteriophage lambda, in the nicking reaction.	cosb is the binding site for terminase, the dna packaging enzyme of ai-12581mbda, and cosn is the adjacent site at which terminase gm-07228es staggered nicks to generate mature lambda dna molecules. there are three binding sites (r3, r2 and r1) within cosb for gpnu1, the small subunit of terminase. a particular transition mutation of r1, known to weaken binding of gpnu1 to r1, has been introduced into the other r sites, and in the present work the effects of r site mutations on nicking of cosn h ...	1993	8254662
formation of linear plasmid multimers promoted by the phage lambda red-system in lon mutants of escherichia coli.	we report here the formation of plasmid linear multimers promoted by the red-system of phage lambda using a multicopy plasmid comprised of lambda red alpha and red beta genes, under the control of the lambda ci857 repressor. our observations have revealed that the multimerization of plasmid dna is dependent on the red beta and reca genes, suggesting a concerted role for these functions in the formation of plasmid multimers. the formation of multimers occurred in a recbcd+ sbcb+ xtha+ lon genetic ...	1993	8254308
high-level expression and purification of human leukotriene a4 hydrolase from insect cells infected with a baculovirus vector.	leukotrienes constitute a group of bioactive compounds derived from arachidonic acid which play important roles in immediate hypersensitivity and inflammation. leukotriene a4 hydrolase (lta4h) is an epoxide hydrolase, catalyzing the hydration of lta4 to ltb4, and also acts an aminopeptidase, with the ability to cleave amides of p-nitroaniline. the cdna for lta4h was cloned using oligonucleotide-directed amplification of the cdna sequence by polymerase chain reaction and by oligonucleotide-based ...	1993	8251746
rapid purification of bacteriophage lambda dna.		1993	8251160
mrna-ribosome interactions.	synthetic mrna analogues were constructed with sequences related to the cro-protein mrna from lambda-phage and prepared by t7 transcription. each mrna contained several thiouridine (thio-u) residues. the regions upstream from the aug initiator codon of the mrna were the same in all the messages, whereas in the downstream part the thio-u residues were placed in selected positions. these positions covered the region from +4 to +16 (a in the initiator aug codon being defined as +1). after binding t ...	1993	8251113
efficient gene transfer with less cytotoxicity by means of cationic multilamellar liposomes.	a simple procedure for the preparation of cationic multilamellar vesicles (mlv) consisting of n-(alpha-trimethylammonioacetyl)-didodecyl-d-glutamate chloride, dilauroyl phosphatidylcholine, and dioleoyl phosphatidylethanolamine in a molar ratio of 1:2:2 was devised. when bacteriophage lambda dna was encapsulated into these liposomes, entrapment efficiency was found to be nearly 100%, and digestibility of the dna was less than 10%. upon encapsulation of the plasmid pch110 into cationic mlv, effic ...	1993	8250862
clinical and molecular evaluation of four patients with partial duplications of the long arm of chromosome 18.	four individuals with partial duplications of the long arm of chromosome 18 were analyzed at the clinical, cytogenetic, and molecular levels. two of the individuals had duplications of the long arm from 18q21.1-qter because of inheritance of an unbalanced translocation. both of these individuals displayed the clinical phenotype characteristic of edwards syndrome. two other patients had de novo interstitial duplications of 18q but did not have a clinical diagnosis of edwards syndrome. the extent ...	1993	8250043
visualization of single molecules of rna polymerase sliding along dna.	transcription requires that rna polymerase binds to promoters buried in nonspecific sites on dna. the search for promoters may be facilitated if the polymerase slides along the molecule of dna. single molecules of escherichia coli rna polymerase were visualized, and their movements on immobilized bacteriophage lambda and t7 dnas were examined. deviating from drifts by bulk flow, about 40 percent of the enzyme molecules moved along the extended dna. the results provide direct evidence for sliding ...	1993	8248804
autoregulation of the escherichia coli heat shock response by the dnak and dnaj heat shock proteins.	all organisms respond to various forms of stress, including heat shock. the heat shock response has been universally conserved from bacteria to humans. in escherichia coli the heat shock response is under the positive transcriptional control of the sigma 32 polypeptide and involves transient acceleration in the rate of synthesis of a few dozen genes. three of the heat shock genes--dnak, dnaj, and grpe--are special because mutations in any one of these lead to constitutive levels of heat shock ge ...	1993	8248205
the escherichia coli hfla locus encodes a putative gtp-binding protein and two membrane proteins, one of which contains a protease-like domain.	the hfla (high frequency of lysogenization) locus of escherichia coli governs the lysis-lysogeny decision of bacteriophage lambda by controlling stability of the phage cii protein. hfla contains three genes, hflx, hflk, and hflc, encoding polypeptides of 50, 46, and 37 kda, respectively. we have determined the nucleotide sequence of 3843 base pairs containing hfla and have found three large open reading frames corresponding to hflx, hflk, and hflc. hflx contains the three sequence motifs typical ...	1993	8248183
cell growth and lambda phage development controlled by the same essential escherichia coli gene, ftsh/hflb.	the lambda phage choice between lysis and lysogeny is influenced by certain host functions in escherichia coli. we found that the frequency of lambda lysogenization is markedly increased in the ftsh1 temperature-sensitive mutant. the ftsh gene, previously shown to code for an essential inner membrane protein with putative atpase activity, is identical to hflb, a gene involved in the stability of the phage cii activator protein. the lysogenic decision controlled by ftsh/hflb is independent of tha ...	1993	8248182
comparison of somatic mutation in a transgenic versus host locus.	somatic mutations can now be quantified in almost any cell type in mice carrying bacterial genes in a lambda phage shuttle vector. mutations induced in vivo are detectable ex vivo, after packaging host-cell dna into phage that are grown on suitable bacteria. however, the transgenic dna differs from many host loci in several ways: it (i) is prokaryotic dna, (ii) is present in multiple tandem copies, and (iii) is heavily methylated and probably not expressed. thus, mutation of a transgene may not ...	1993	8248160
expression and biochemical properties of a protein serine/threonine phosphatase encoded by bacteriophage lambda.	the predicted amino acid sequence encoded by the open reading frame 221 (orf221) of bacteriophage lambda exhibited a high degree of similarity to the catalytic subunits of a variety of protein serine/threonine phosphatases belonging to pp1, pp2a, and pp2b groups. cloning and expression of the orf221 gene in escherichia coli provided direct evidence that the gene codes for a protein serine/threonine phosphatase. the single-subunit recombinant enzyme was purified in soluble form and shown to posse ...	1993	8248155
[study of the conformational and intradynamic changes of phage lambda dna molecules by laser correlation spectroscopy].	conformational changes of lambda phage dna macromolecules were investigated by dynamic light scattering (dls) method. it was shown that the persistence length of b-form dna in some conditions could could decline to approximately 12 nm. when studying the internal motions in the linear form of phage dna macromolecule, a deviation was observed of the experimental angular dependence of the first cumulant gamma from theoretical predictions for the gamma in rouse-zimm model including hydrodynamic inte ...	1993	8246936
[cloning of the gene for thermostable thermus aquaticus yt1 dna polymerase and its expression in escherichia coli].	using the phasmid vector psl5, the genomic dna fragment of t. aquaticus yt1 which contained the thermostable dna polymerase (taq-polymerase) gene was cloned. the bglii fragment of this genome locus was subcloned in the bamhi site of the puc19 plasmid. to optimize the taq-polymerase gene expression in e. coli cells, the gene was cloned in the correct reading frame regarding the initiation atg codon of the ppr-tgatg-1 expression vector. the gene expression in this vector was controlled by the phag ...	1993	8246933
antigen-dependent stimulation by bone marrow-derived mast cells of mhc class ii-restricted t cell hybridoma.	this paper describes a new role for mast cells as being able to present ag to immune t cells. a mouse bone marrow-derived mast cell population obtained after 3 wk of culture in a conditioned medium has been shown to express a variety of membrane-associated ag, including mhc class ii and class i ag, cd23, cd32, high affinity receptor for ige, and cd4. expression of mhc class ii molecules was up-regulated upon stimulation with lps but not with ifn-gamma and was down-regulated after exposure of mas ...	1993	8245470
energy-dependent degradation of lambda o protein in escherichia coli.	protein o of bacteriophage lambda is a short-lived protein which has a key role in the replication of the phage dna in escherichia coli. here we present evidence that lambda o degradation is energy dependent: it is impaired by cyanide and alpha-methylglucoside, both of which inhibit cellular energy metabolism. removal of these inhibitors restored the degradation of lambda o. our experiments suggest that limited amounts of cellular energy are sufficient to support lambda o degradation. in additio ...	1993	8244945
in vivo mutagenesis induced by cc-1065 and adozelesin dna alkylation in a transgenic mouse model.	although considerable work has focused on characterizing the bonding chemistry and sequence selective alkylation of dna by cyclopropylpyrroloindole compounds, little is known about the molecular consequence of their n-3-adenine adducts in whole animal systems. we have utilized a transgenic mouse system, harboring a lambda phage shuttle vector, to assess the mutagenic potential of the antitumor compounds cc-1065 and adozelesin and, for the first time, to track the in vivo fate of their unique dna ...	1993	8242625
detection of mycoplasma meleagridis by polymerase chain reaction.	a pair of 32 base primers was synthesized based on the dna sequence data of a mycoplasma meleagridis (mm) species-specific recombinant, pmm-2. the primers were used in a mm-polymerase chain reaction (pcr) to amplify a target dna of approximately 850 bp. annealing temperatures ranging from 58 degrees c to 61 degrees c could be used for the mm-pcr without loss of specificity. the primers amplified 1 ng of dna from 17 strains of mm, but not 10 ng of dna from 16 heterologous species of avian mycopla ...	1993	8236783
posttranscriptional control of the lysogenic pathway in bacteriophage lambda.		1993	8234786
use of a gene encoding a suppressor trna as a reporter of transcription: analyzing the action of the nun protein of bacteriophage hk022.	the nun protein of phage hk022 blocks the expression of genes that lie downstream of the nut sites of phage lambda. nun is believed to act by promoting premature termination of transcription at or near these sites. to test this hypothesis and to facilitate mapping the sites of termination, we inserted a gene encoding a suppressor trna immediately downstream of the lambda nutl site and determined the effect of nun on trna level. we found that nun severely reduced the accumulation of mature, biolo ...	1993	8234323
bacterially expressed fabs of monoclonal antibodies neutralizing tumour necrosis factor alpha in vitro retain full binding and biological activity.	antibody fragments specific for the human tumour necrosis factor alpha (tnf alpha) have been cloned from lambda combinatorial expression libraries using total rna obtained from three different hybridoma cell lines of therapeutic interest. the previously described bacteriophage lambda vectors, lambda hc2 and lambda lc1, were modified to create unique antibody cloning sites in the combinatorial construct and a novel tag peptide was inserted at the c-terminal end of the expressed fd chain. sequence ...	1993	8232337
targeted mutagenesis of simian virus 40 dna mediated by a triple helix-forming oligonucleotide.	triple-helical dna can be formed by oligonucleotides that bind as third strands of dna in a sequence-specific manner in the major groove in homopurine/homopyrimidine stretches in duplex dna. such triple helix-forming oligonucleotides have been used to inhibit gene expression by blocking transcription factor access to promoter sites in transient expression assays. in an alternative approach to genetic manipulation using triplex dna, we show that triplex-forming oligonucleotides can be used to pro ...	1993	8230456
a programmed translational frameshift is required for the synthesis of a bacteriophage lambda tail assembly protein.	two proteins, one of 31 kda and one of 16 kda, are encoded by a segment of the phage lambda tail gene region that contains two overlapping reading frames, neither of which is long enough to encode the larger protein. we show that the abundant 16-kda protein (gpg) is encoded by the upstream open reading frame, gene g. the 31-kda protein, gpg-t, is encoded jointly by gene g and the overlapping downstream t open reading frame. gpg-t is synthesized as the result of a translational frameshift that oc ...	1993	8230192
function of the grpe heat shock protein in bidirectional unwinding and replication from the origin of phage lambda.	the initiation of dna replication by phage lambda depends on a specialized nucleoprotein structure that provides for the precise localization and activity of the escherichia coli dnab helicase at the lambda replication origin. previous work has shown that the dnaj and dnak heat shock proteins function in the initiation pathway by releasing the dnab helicase from the initiation complex to carry out localized unwinding of origin dna. this dnaj.dnak pathway results in mainly unidirectional dna unwi ...	1993	8227083
isolation and characterization of clpx, a new atp-dependent specificity component of the clp protease of escherichia coli.	we have used 14c-labeled bacteriophage lambda o-dna replication protein as a probe to identify and purify escherichia coli proteases capable of its degradation. in this manner, five different proteases (termed lop) have been identified capable of degrading lambda o protein to acid-soluble fragments in an atp-dependent fashion. one of these activities was purified to homogeneity and shown to be composed of two different polypeptides. the 23,000-da component (lopp) was identified as the previously ...	1993	8226769
a novel phage lambda replacement cre-lox vector that has automatic subcloning capabilities.	we have developed a novel phage lambda replacement cloning vector, lambda pan. lambda pan allows one to automatically subclone the insert as a plasmid using the cre-loxp site-specific recombination system. this eliminates the need to subclone insert fragments and permits the rapid structural analysis of insert dna. lambda pan is similar to other phage lambda replacement vectors taking inserts ranging in size from 5 to 19 kb. we have placed the pyrg gene of aspergillus nidulans on the vector as a ...	1993	8224901
cloning, purification and characterization of the bseci dna methyltransferase from bacillus stearothermophilus.	the gene (bsecim) encoding the bseci dna methyltransferase (mtase; m.bseci) from a bacillus stearothermophilus species was cloned and expressed in escherichia coli using plasmid vector pbr322. selection of transformants carrying bsecim was based on the resistance of the modified plasmid to cleavage by bseci. the mtase was purified to homogeneity and further characterized. its size as determined by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis and size exclusion chromatography was 68 ...	1993	8224900
synthesis of two bacteriophage lambda s proteins in an in vivo system.	bacteriophage lambda has two genes which are essential for lysis: r, a gene encoding a 158-amino-acid (aa) transglycosylase that attacks the peptidoglycan, and s, a gene encoding two inner-membrane-associating proteins, designated s105 and s107 for their predicted lengths in aa residues. s105 and s107 are thought to have opposing roles in lysis, with the former acting as the lethal lysis effector and the latter as a lysis inhibitor. here, we used a t7-polymerase-mediated expression system to sho ...	1993	8224899
esterase from the oil-degrading acinetobacter lwoffii rag-1: sequence analysis and over-expression in escherichia coli.	the est gene encoding an esterase from acinetobacter lwoffii rag-1 was cloned into e. coli under the control of the pl promoter of the phage lambda. the n-terminal sequence of the first 20 amino acids of the heterologous expressed esterase corresponded to that obtained from the nucleotide sequence. antibodies prepared against the over-expressed recombinant esterase in e. coli were used to locate the enzyme primarily in the membrane fractions of a. lwoffii rag-1. comparison with homologous protei ...	1993	8224790
the human immunoglobulin kappa locus. characterization of the partially duplicated l regions.	the l regions are parts of the c kappa proximal (p) and distal (d) copies of the human immunoglobulin kappa locus and are therefore called the lp and ld regions. the two regions with their 25 v kappa genes and pseudogenes have now been cloned, thus completing the cloning of the kappa locus. lp has been linked to the neighboring ap and b regions, while ld was linked to ad. there is good evidence that at the other side of ld, i.e. towards the centromere, the end of the locus has been reached. most ...	1993	8223862
mutagenicity of methyl methanesulfonate (mms) in vivo at the dlb-1 native locus and a laci transgene.	methyl methanesulfonate (mms) is an extraordinarily poor mutagen compared to ethylnitrosourea (enu) or even x-rays. in lung fibroblasts in vivo, mms has been shown to induce many micronuclei but few, if any, mutations at the hpt locus. we wondered if the lack of mutations might be due to the lack of division and dna synthesis in fibroblasts in vivo, which would permit substantial time for differential repair of dna lesions. this idea was tested in the small intestine, a tissue in which the cells ...	1993	8223513
complementation of bacteriophage lambda integrase mutants: evidence for an intersubunit active site.	site-specific recombination of bacteriophage lambda starts with the formation of higher-order protein--dna complexes, called 'intasomes', and is followed by a series of steps, including the initial dna cleavage, top-strand exchange, branch migration and bottom-strand exchange, to produce recombinant products. one of the intasomes formed during excisive recombination (the attl complex) is composed of the phage-encoded integrase (int), integration host factor (ihf) and one of the recombination sub ...	1993	8223467
synaptic intermediates in bacteriophage lambda site-specific recombination: integrase can align pairs of attachment sites.	bacteriophage lambda uses site-specific recombination to move its dna into and out of the escherichia coli genome. the recombination event is mediated by the recombinase integrase (int) together with several accessory proteins through short specific dna sequences known as attachment sites. a gel mobility shift assay has been used to show that, in the absence of accessory proteins, int can align and hold together two dna molecules, each with an attachment site, to form stable non-covalent 'bimole ...	1993	8223466
rna-mediated specificity of dna packaging into hybrid lambda/phi 29 proheads.	a small rna (prna, 174 nt) is known to be essential for dna packaging in bacteriophage phi 29. however, in an in vitro dna packaging system based on hybrid lambda/phi 29 proheads (made up of head proteins from phage lambda and connectors from phage phi 29), the specificity of dna packaging is lost, and different rna molecules fulfil the requirements for dna packaging, albeit with less efficiency than phi 29 prna. competition assays with rnas from different sources have shown that phi 29 connecto ...	1993	8223455
mapping of the gene coding for a paraneoplastic encephalomyelitis antigen (hud) to human chromosome site 1p34.	a cdna clone coding for a human brain protein (hud), which is highly homologous to the drosophila neuronal determination protein elav and elicits antibody formation in a high proportion of patients with paraneoplastic encephalomyelitis, was used to isolate a lambda phage recombinant clone, including a large fragment of the relevant human genomic region. the fragment proved to be an efficient probe for the precise subregional mapping of the gene by molecular in situ hybridization onto extended hu ...	1994	8222755
molecular cloning, characterization and expression of mn-superoxide dismutase from the rubber tree (hevea brasiliensis).	a genomic clone encoding manganese-containing superoxide dismutase (sod; ec 1.15.1.1) was isolated from a hevea brasiliensis genomic library made in lambda phage embl3 by using a heterologous cdna probe of mnsod from nicotiana plumbaginifolia. the nucleotide sequence of 4968 bp from the genomic clone was determined. based on the putative translation initiation codon and stop codon, pcr primers were designed and utilized for cloning the full-length cdna from total mrna. of the two distinct cdnas ...	1993	8219064
kinetic characterization of the atpase activity of the dna packaging enzyme from bacteriophage lambda.	terminases are enzymes common to all of the complex double-stranded dna viruses and are required for viral assembly. these enzymes function to excise a single viral genome from a concatemeric dna precursor and package it into a preformed protective protein shell or capsid. atp hydrolysis by these enzymes has been described and appears to be critical to the packaging process. we have previously characterized the endonuclease activity of purified terminase from bacteriophage lambda [tomka, m. a., ...	1993	8218275
characterization of a restriction barrier and electrotransformation of the cyanobacterium nostoc pcc 7121.	we have investigated host restriction as a barrier to transformation and developed a method for gene transfer into the previously untransformable, heterotrophic cyanobacterium nostoc pcc 7121. a restriction endonuclease, designated nsp 7121i, has been partially purified by phosphocellulose chromatography of nostoc cell extracts. comparisons of nsp 7121i digests of bacteriophage lambda and plasmid dnas with computer-generated restriction fragment profiles showed that nsp 7121i is an isoschizomer ...	1993	8215799
dynamic properties of nucleic acids in biosupramolecular systems, as studied by 31p nmr.	the dynamic properties of nucleic acids in five different types of intact supramolecular systems, namely, chicken erythrocyte chromatin, the wild type and a deletion mutant of the lambda phage, lipid-containing phage pm2, and alteromonas espejiana ribosomes, were investigated by means of 31p solid-state nmr. the nucleic acids in the different supramolecular systems showed unique dynamic properties, which are closely connected with their functions. the total anisotropy of the phosphorus chemical ...	1994	8206876
structure of the bovine lactoferrin-encoding gene and its promoter.	lactoferrin (lf), a ferric ion (fe3+)-binding glycoprotein, is found most notably in milk, probably to mediate protection against microbial infection of the mammary gland. based on an initial isolation and sequencing of a complete cdna of the bovine lf gene (blf), the complete gene was obtained from genomic libraries on five overlapping phage lambda embl3 clones. a detailed restriction map and the complete exon/intron structure of the gene are presented, together with 1 kb of sequence data of th ...	1994	8206385
the 9-kda calbindin gene of rousettus aegyptiacus: its identification and isolation from a genomic library.	a genomic library of the fruit bat (rousettus aegyptiacus) was constructed in lambda phage gt11. the titre of the library was determined to be 2 x 10(5) pfu/ml. the genomic library was amplified and the titre of the amplified library increased 300-fold to 7 x 10(7) pfu/ml. the library was screened by in situ hybridization techniques using a fragment of the mouse 9-kda calbindin cdna as a probe. screening of 10(5) plaques yielded a positive clone. three additional rounds of screening were perform ...	1994	8205389
restriction mapping of phage lambda vectors using non-radioactive methods.	in order to take advantage of non-radioactive methods, we have developed two plasmids (p lambda le and p lambda re) for mapping restriction sites of long inserts cloned in phage lambda vectors. these plasmids are constructed by cloning the left 402-bp and right 560-bp phage lambda genome ends, respectively. to map restriction sites, the cloned sequences in p lambda le and p lambda re are labeled with digoxygenin and hybridized to partially digested lambda dna. the ladder of bands detected with t ...	1994	8200536
molecular cloning of the gene encoding the mouse parathyroid hormone/parathyroid hormone-related peptide receptor.	the parathyroid hormone/parathyroid hormone-related peptide receptor (pthr) is a g-protein-coupled receptor containing seven predicted transmembrane domains. we have isolated and characterized recombinant bacteriophage lambda embl3 genomic clones containing the mouse pthr gene, including 10 kilobases of the promoter region. the gene spans > 32 kilobases and is divided into 15 exons, 8 of which contain the transmembrane domains. the pthr exons containing the predicted membrane-spanning domains ar ...	1994	8197183
structure of the human na+/glucose cotransporter gene sglt1.	intestinal uptake of dietary glucose and galactose is mediated by the sglt1 na+/glucose cotransporter of the brush border. an sglt1 missense mutation underlies hereditary glucose/galactose malabsorption, characterized by potentially fatal diarrhea; conversely, oral rehydration therapy exploits normal transport to alleviate life-threatening diarrhea of infectious origin. we have mapped the entire human sglt1 na+/glucose cotransporter gene from cosmid and lambda phage clones representing a genomic ...	1994	8195156
a role for residue 151 of lamb in bacteriophage lambda adsorption: possible steric effect of amino acid substitutions.	lamb is the cell surface receptor for bacteriophage lambda. lamb missense mutations yielding resistance to lambda have been previously grouped in two classes. class i mutants block growth of lambda with wild-type host range (lambda h+) but support growth of one-step extended-host-range mutants (lambda h). class ii mutants block lambda h but support growth of two-step extended host range mutants (lambda hh*). while class i mutations occur at 11 different amino acid sites, in five distinct portion ...	1994	8195074
cloning of a streptococcus sobrinus oligo-isomaltosaccharide synthase gene and characterization of its product.	a streptococcus sobrinus gene coding for a glucosyltransferase (gtf)-s was cloned into escherichia coli, using the bacteriophage lambda l47.1 and the plasmid vector pacyc184. the md124 clone obtained expressed a 155 kda gtf-s which did not react with any antisera against gtf-s1, -s2 and -i enzymes. the recombinant enzyme (designated rgtf-s3) was homogeneously purified from the md124 cell-extract and characterized. the purified rgtf-s3 synthesized primer-independently alpha-1,6-linked linear olig ...	1993	8193601
determining the dna sequence elements required for binding integration host factor to two different target sites.	binding sites for the escherichia coli protein integration host factor (ihf) include a set of conserved bases that can be summarized by the consensus sequence watcaannnnttr (w is da or dt, r is da or dg, and n is any nucleotide). however, additional 5'-proximal bases, whose common feature is a high da+dt content, are also thought to be required for binding at some sites. we examine the relative contribution of these two sequence elements to ihf binding to the h' and h1 sites in attp of bacteriop ...	1994	8188600