Publications

analysis of spontaneous and induced mutations in transgenic mice using a lambda zap/laci shuttle vector.	a short term, in vivo mutagenesis assay has been developed utilizing a lacl target gene contained within a lambda zap shuttle vector which has been incorporated into transgenic mice. following chemical exposure, the target gene was recovered from mouse genomic dna by mixing the dna with in vitro lambda phage packaging extract. mutations within the lacl target were identified by infecting host e. coli with the packaged phage and plating on indicator plates containing xgal. phage plaques with muta ...	1991	1836179
rna polymerase bound to the pr promoter of bacteriophage lambda inhibits open complex formation at the divergently transcribed prm promoter. implications for an indirect mechanism of transcriptional activation by lambda repressor.	we demonstrate that rna polymerase bound at the pr promoter of bacteriophage lambda can repress transcription initiation from the divergently transcribed prm promoter in vitro. using abortive initiation and run-off transcription experiments we show that inactivating mutations introduced into either the -10 or -35 regions of pr result in a significant increase in the rate of formation of transcriptionally competent complexes at the prm promoter. this is due primarily to an increase in the rate co ...	1991	1836235
chain bias in chi-stimulated heteroduplex patches in the lambda ren gene is determined by the orientation of lambda cos.	heteroduplex patch recombinants have received information in one dna chain but have not recombined flanking markers. evidence regarding which chain is exchanged bears on the structure of recombination intermediates. the direction of travel along dna of recbcd recombinase, the central enzyme in the escherichia coli recbcd pathway of homologous recombination, is determined in phage lambda by the orientation of the packaging origin, cos. cos is a double-chain cut site which serves as a preferred en ...	1991	1836442
[a universal instinct for designing thermoregulated promotors in gram-positive bacteria].	the construction of plasmid pvkh300, which is useful for modifying any promoter into the thermoregulated form in b. subtilis cells, is presented. the main features of the plasmid are the presence of effectively expressed in b. subtilis lambda c1857 gene and recognition site of bglii restriction enzyme between or2 and or3 lambda phage operator sites. promoterless alpha-amylase gene of b. amyloliquefaciens is used as a reporter gene for promoter cloning into bglii site of pvkh300. examples of prom ...	1991	1836528
[trigger effect in switching the phage lambda genome].	the paper presents a mathematical model of the trigger switching of lambda phage genome on the basis of the molecular ideas by ptashne. the sigmoid character of the gene cro promotor was explained via lambda-repressor concentration. the temperature switching of genetic trigger was attributed to the existence of permissive and restrictive temperatures by means of the fokker-plank probability density formalism. the dependence of the lambda repressor average concentration of the uv-radiation dose w ...	1991	1836529
[synthesis, secretion, and proteolytic degradation of diphtheria toxin in escherichia coli].	the recombinant plasmids have been constructed encoding the synthesis of a full-sized diphtheria toxin from its own or pr, pl-promoters of bacteriophage lambda in escherichia coli cells. the high level constitutive synthesis of toxin results in slow cell growth and plasmid elimination. the toxin was mainly detected in the periplasm, partially in the membrane and to a less extent in the cytoplasm and culturing medium. the dimeric form of toxin was found in the cytoplasm. participation of toxin b- ...	1991	1836836
development of genomic probes to sarcocystis cruzi (apicomplexa).	a genomic library of sarcocystis cruzi sporozoite dna was constructed in bacteriophage lambda gt10. recombinant phages containing insert dna were selected by growth on escherichia coli strain c600 hfla150. of 14 clones examined, 11 contained dna inserts ranging in size from approximately 1.45 kilobase (kb) to 6.18 kb. insert dna from four of these clones specifically hybridized to 32p-labelled s. cruzi merozoite dna. one of these insert dna, clone sl41, was selected and labelled with 32p. this p ...	1991	1837193
the murine mov-34 gene: full-length cdna and genomic organization.	the mov-34 mutation is a recessive embryonic lethal mutation caused by experimental introduction of a recombinant moloney murine leukemia provirus into the mouse germline. we have cloned a full-length cdna from the mov-34 gene, the transcription unit disrupted by the proviral integration. this cdna is predicted to encode a novel 321-amino acid, 36-kda protein of unknown function. overlapping phage lambda clones containing the entire mov-34 gene have been isolated. the mov-34 gene spans just over ...	1991	1837787
the kila operon of promiscuous plasmid rk2: the use of a transducing phage (lambda pklaa-1) to determine the effects of the lethal klaa gene on escherichia coli cells.	the kil-kor regulon of promiscuous plasmid rk2 includes the replication initiator gene trfa and several potentially host-lethal kil loci (kila, kilb, kilc, kile), whose functions may be involved in plasmid maintenance or broad host range. the kila locus consists of a single operon of three genes (klaa, klab, klac), each of which is lethal when expressed from the klaa promoter in the absence of repressors encoded by kora and korb. in this study, we examined the effects of the unregulated klaa gen ...	1991	1838127
recombinagenic processing of uv-light photoproducts in nonreplicating phage dna by the escherichia coli methyl-directed mismatch repair system.	nonreplicating lambda phage dna in homoimmune escherichia coli lysogens provides a useful model system for study of processes that activate dna for homologous recombination. we measured recombination by extracting phage dna from infected cells, using it to transfect reca recipient cells, and scoring the frequency of recombinant infective centers. with unirradiated phage, recombinant frequencies were less than 0.1%. however, recombination could be increased over 300-fold by prior uv irradiation o ...	1991	1838344
molecular characterization of a cluster of at least two glucosyltransferase genes in streptococcus salivarius atcc 25975.	the oral micro-organism streptococcus salivarius atcc 25975 synthesizes extracellular glucosyltransferases (gtfs) which polymerize the glucose moiety of sucrose into glucan polymers. two separate genes encoding the activities of a gtf-i (a gtf that synthesizes an insoluble product) and a gtf-s (a gtf that synthesizes soluble product) were cloned into bacteriophage lambda l47.1. the inserts in the lambda-clones were characterized by restriction mapping and southern hybridization and were found to ...	1991	1838391
[complexity analysis of a genome. ii. extensive homology zones in bacteriophage lambda].	the suggested earlier complexity approach for detecting structural regularities in primary structures of nucleic acids is illustrated by using lambda phage as an example. among the most interesting regularities detected in the lambda phage genome are the following: (a) the presence of "extended homology zones" i.e. fragments in which block transpositions of duplicative type predominate explicitly; (b) the abundance of palindrome-hairpin structures and duplications in the origins and termini of m ...	1991	1839056
role of the escherichia coli grpe heat shock protein in the initiation of bacteriophage lambda dna replication.	initiation of replication of lambda dna requires assembly of the proper nucleoprotein complex consisting of the lambda origin of replication-lambda o-lambda p-dnab proteins. the dnaj, dnak and grpe heat shock proteins destabilize the lambda p-dnab interaction in this complex permitting dnab helicase to unwind lambda dna near ori lambda sequence. first step of this disassembling reaction is the binding of dnak protein to lambda p protein. in this report we examined the influence of dnaj and grpe ...	1991	1839120
rnaselll activation of bacteriophage lambda n synthesis.	the bacteriophage lambda n gene product is one of the first genes expressed during phage development. n protein allows the expression of other phage genes by altering the transcription elongation process so as to prevent transcription termination. we have found that n levels may be modulated soon after induction or infection. using n-lacz fusions, we determined that cells containing rnaselll have at least a fourfold greater expression than cells defective for rnaselll. this effect is exerted at ...	1991	1839745
a probabilistic model for genetic recombination of nonreplicating lambda-phage dna, stimulated by "mismatch repair" of uv photoproducts.	genetic recombination of nonreplicating phage lambda-dna, during infection of homoimmune lysogenic bacteria, was previously observed to be dramatically stimulated by prior uv irradiation of the phages, even when the escherichia coli hosts lacked the major uv-photo-product excision-repair system (uvrabc). uvrabc-independent recombination of circular phage molecules depends on host muthls functions and on undermethylation of adenines at gatc sites in the phage dna, and thus appears to be the resul ...	1991	1839959
selection of lacz operon fusions in genes of gluconate metabolism in e. coli. characterization of a gntt::lacz fusion.	the initial steps involved in the utilization of gluconate by e. coli, its incorporation into the cell and subsequent phosphorylation to gluconate 6-phosphate, conform two systems that duplicate activities. these systems, gnti and gntii, are specified by two sets of genes distinctly regulated and located respectively at the mala-asd (75 min) and fdp-vals (96 min) regions of the bacterial chromosome. the presence of duplicate activities in the metabolism of gluconate of e. coli, has made difficul ...	1991	1843569
a salmonella typhimurium virulence protein is similar to a yersinia enterocolitica invasion protein and a bacteriophage lambda outer membrane protein.	the phop-phoq-regulated pagc locus is essential for full virulence and survival within macrophages of salmonella typhimurium. the protein product, dna sequence, and transcript of pagc were determined. the pagc locus encodes a single 188-amino-acid membrane protein that is similar to the ail-encoded eucaryotic cell invasion protein of yersinia enterocolitica and the lom-encoded protein of bacteriophage lambda. the similarity of pagc and ail to lom leads us to hypothesize that lom is a virulence p ...	1991	1846140
transposon tn5supf-based reverse genetic method for mutational analysis of escherichia coli with dnas cloned in lambda phage.	an efficient method for systematic mutational analysis of the escherichia coli genome was developed. it entails tn5supf transposition to lambda-e. coli hybrid phage clones (kohara library) and then transduction of recipient cells to sup+. essential and nonessential genes are distinguished by the ability of insertion mutant phage to form haploid versus only heterozygous partial diploid bacterial recombinants.	1991	1846153
t cell receptor alpha and beta gene expression in a murine antigen-specific t suppressor lymphocyte clone with cytolytic potential.	the composition of alpha and beta tcr genes was analyzed in a murine bsa-specific ts cell clone with cytolytic potential. the isolated poly(a)+ mrna from ts cell clone bvi/5 was used to construct a cdna library in the bacteriophage lambda gt11. full-length cdna clones specific for tcr alpha and tcr beta genes have been detected and isolated by hybridization with specific oligonucleotide probes. the functional rearranged tcr alpha gene is composed of a member of the v alpha 1 family, the junction ...	1991	1846162
two-step cloning and expression in escherichia coli of the dna restriction-modification system stylti of salmonella typhimurium.	the stylti restriction-modification system is common to most strains of the genus salmonella, including salmonella typhimurium. we report here the two-step cloning of the genes controlling the stylti system. the stylti methylase gene (mod) was cloned first. then, the companion endonuclease gene (res) was introduced on a compatible vector. a strain of s. typhimurium sensitive to the coliphage lambda was constructed and used to select self-modifying recombinant phages from a res- mod+ s. typhimuri ...	1991	1846861
bacteriophage lambda promoters pl and pr: sequence determinants of in vivo activity and of sensitivity to the dna gyrase inhibitor, coumermycin.	sequence encompassing the region between bp -43 and +8 of the pl and pr promoters of bacteriophage lambda, as well as sequence variants of these promoters, were compared with respect to their ability to drive a promoterless cat gene in vivo. for both pl- and pr-based promoters, variants with one nonconsensus bp rather than the consensus promoter were found to be maximally active. determination of promoter function in csh26 and c600 revealed a marked strain dependence in the activity of some prom ...	1991	1847348
analysis of the escherichia coli nusa10(cs) allele: relating nucleotide changes to phenotypes.	the escherichia coli nusa gene product, known to influence transcription elongation, is essential for both bacterial viability and growth of lambdoid phages. we report the cloning and sequencing of the conditionally lethal nusa10(cs) allele. changes from nusa+ were observed at nucleotides 311 and 634. functional studies showed that both nucleotide changes are necessary for the cold-sensitive phenotype, although bacteria with the change at 634 grew more slowly at 30 degrees c than the nusa+ contr ...	1991	1847364
analysis of an escherichia coli dnab temperature-sensitive insertion mutation and its cold-sensitive extragenic suppressor.	an escherichia coli mutant, ts121, was isolated following random insertional mutagenesis using phage lambda mu transposition. the mutant phenotype includes inability to form colonies at temperatures above 38 degrees c and inability to propagate phage lambda at all temperatures. a lambda i434 ci- (ts121)+ transducing phage was isolated on the basis of its ability to form plaques on ts121 mutant bacteria. using this transducing phage, it was shown through complementation and protein analyses, that ...	1991	1847383
lambda yes: a multifunctional cdna expression vector for the isolation of genes by complementation of yeast and escherichia coli mutations.	this work describes a multifunctional phage lambda expression vector system, lambda yes, designed to facilitate gene isolation from eukaryotes by complementation of escherichia coli and saccharomyces cerevisiae mutations. lambda yes vectors have a selection for cdna inserts using an oligo adaptor strategy and are capable of expressing genes in both e. coli and s. cerevisiae. they also allow conversion from phage lambda to plasmid clones by using the cre-lox site-specific recombination system, re ...	1991	1848010
the ubiquitin-encoding multigene family of flax, linum usitatissimum.	ubiquitin (ubq), a 76-amino acid (aa) protein, is found in all eukaryotic organisms and is one of the most conserved proteins so far studied. it is implicated in many cellular processes. the ubq-encoding genes (ubq) are generally present as a multigene family. in flax, we have estimated that this multigene family contains at the most ten members. the initial flax ubq sequences were isolated from a flax genomic library in lambda embl4 using a heterologous arabidopsis thaliana ubq probe. an 916-bp ...	1991	1850710
[a comparative study of the functions of recbcd-nuclease from escherichia coli and "recombinase", encoded by plasmid r1drd-19].	the recbcd nuclease of escherichia coli and "recombinase" determined by r1drd-19 plasmid (the latter is able to replace at least partially the indicated cellular enzyme) were shown to differ from each other in some essential features. the product encoded by the plasmid as distinct from recbcd nuclease practically is not sensitive to inhibition by gams protein of the lambda phage. earlier, it was found that the presence of r1drd-19 plasmid in the recbc cells restores the level of the total atp-de ...	1991	1851536
cloning, sequencing, and expression of the gene coding for bile acid 7 alpha-hydroxysteroid dehydrogenase from eubacterium sp. strain vpi 12708.	southern blot analysis indicated that the gene encoding the constitutive, nadp-linked bile acid 7 alpha-hydroxysteroid dehydrogenase of eubacterium sp. strain vpi 12708 was located on a 6.5-kb ecori fragment of the chromosomal dna. this fragment was cloned into bacteriophage lambda gt11, and a 2.9-kb piece of this insert was subcloned into puc19, yielding the recombinant plasmid pbh51. dna sequence analysis of the 7 alpha-hydroxysteroid dehydrogenase gene in pbh51 revealed a 798-bp open reading ...	1991	1856160
human type vii collagen: cdna cloning and chromosomal mapping of the gene.	a human keratinocyte cdna expression library in bacteriophage lambda gt11 was screened with the purified igg fraction of serum from a patient with epidermolysis bullosa acquisita, which had a high titer of anti-type vii collagen antibodies. screening of approximately 3 x 10(5) plaques identified 8 positive clones, the largest one (k-131) being approximately 1.9 kilobases in size. dideoxynucleotide sequencing of k-131 indicated that it consisted of 1875 base pairs and contained an open reading fr ...	1991	1871109
the porcine tumor necrosis factor-encoding genes: sequence and comparative analysis.	we have cloned and sequenced a 10.22-kb fragment of the genomic locus of the porcine tumor necrosis factor-encoding genes, tnf-alpha and tnf-beta. a liver genomic dna library, partially digested with sau3ai, was cloned into the phage lambda embl4 and screened with a porcine tnf-alpha cdna probe. analysis showed that both the tnf-alpha and tnf-beta genes were present on the cloned fragment. in addition, the cloned fragment contained about 2 kb of repetitive sequences 5' to the tnf-beta gene. the ...	1991	1874444
small acidic peptides are bound to e. coli dna.	low molecular weight peptides have been isolated by alkali extraction from deproteinized dna of e. coli cells grown in the presence of radioactive glutamic acid or orthophosphate. the labeled peptides, purified by gel filtration chromatography on sephadex g25 and g10, contain prevailingly glutamic acid, aspartic acid, glycine, serine and alanine. electrophoretic studies at different ph show that some peptide fractions contain a phosphoric residue. the n-terminus of the phosphorylated peptides is ...	1991	1875921
a copy of exon 3-intron 3 from the barley aleurain gene is present on chromosome 2.	a genomic clone from hordeum vulgare l. cv. himalaya contains 700 bp of dna that is homologous with a high degree of nucleotide sequence similarity to exon 3-intron 3 from the gene for the thiol protease, aleurain. genomic southern blot mapping data indicate that this clone in phage lambda had not undergone rearrangement, and no other sequences homologous to aleurain are present on it. although exon 3 in aleurain encodes the polypeptide region cleaved during proteolytic processing of the proenzy ...	1991	1884002
quantitative study of protein association at picomolar concentrations: the lambda phage cl repressor.	a method has been developed for radiolabeling the lambda cl repressor to a specific activity sufficiently high to permit accurate quantitation of the protein in the picomolar range of concentration. procedures are described whereby the labeled protein can be used for accurate quantitative study of the energetics of repressor assembly by large zone analytical gel chromatography. this methodology is applicable to other systems in which the stoichiometry and energetics of tightly associating dna bi ...	1991	1888038
schistosoma haematobium and s. japonicum: analysis of the ribosomal rna genes and determination of the "gap" boundaries and sequences.	we have determined the intragenic organization of the rrna genes of schistosoma haematobium and s. japonicum and found them to be similar to that of s. mansoni and other eukaryotes. an entire ribosomal repeat approximately 10 kbp in size from each species was isolated as a sali fragment from a genomic library constructed in bacteriophage lambda. the segments encoding both the small and large rrnas have been identified using three cloned ecori fragments of s. mansoni as probes. there were three e ...	1991	1889469
intersubunit disulfide-bonded lambda-cro protein.	site-directed mutagenesis has been employed to substitute cysteine for valine at position 55, which is located on the dimer interface of the cro protein of bacteriophage lambda. it has been found that the cys55 cro protein (cro vc55) spontaneously forms a stable disulfide-bonded dimer in the absence of a reducing agent. uv-cd and nmr data showed that the mutant protein retains the conformation of the wild cro protein and has acquired significant heat-stability. however, its specific dna-binding ...	1991	1891462
isolation of bradyrhizobium japonicum dna sequences that are transcribed at high levels in bacteroids.	dna sequences have been isolated that are expressed at high levels in bacteroids, the differentiated form of the soybean microsymbiont, bradyrhizobium japonicum. random-primed cdna was synthesized using total rna isolated from purified b. japonicum bacteroids or from cells grown in culture. when used directly to screen bacteriophage lambda libraries, these cdna probes produced a high background hybridization signal due to sequence similarity between b. japonicum and e. coli ribosomal dna (rdna) ...	1991	1896009
rna polymerases from pseudomonas aeruginosa and pseudomonas syringae respond to escherichia coli activator proteins.	the activities of rna polymerases (rnaps) from pseudomonas aeruginosa and pseudomonas syringae were compared with that of escherichia coli rnap. all three enzymes are able to initiate transcription at the trpba promoter of p. aeruginosa and at the coliphage lambda promoters, prm and pre, in response to heterospecific activators (trpi protein, repressor, and cii protein, respectively). however, both pseudomonas polymerases have less stringent requirements for promoter recognition in the absence o ...	1991	1898924
bacillus subtilis dna polymerase iii: complete sequence, overexpression, and characterization of the polc gene.	genomic dna encompassing polc, the structural gene specifying bacillus subtilis dna polymerase iii (poliii), was sequenced and found to contain a 4311-bp open reading frame (orf) encoding a 162.4-kda polypeptide of 1437 amino acids (aa). the orf was engineered into an escherichia coli expression plasmid under the control of the coliphage lambda repressor. derepression of e. coli transformants carrying the recombinant vector resulted in the high-level synthesis of a recombinant dna polymerase ind ...	1991	1901559
cloning and expression in cos-1 cells of a full-length cdna encoding human coagulation factor x.	a 1.5-kb cdna (fx) encoding full-length human coagulation factor x was isolated from a human fetal liver cdna library. the identity of the insert in a selected phage lambda clone was confirmed to be fx by nucleotide (nt) sequence analysis and restriction mapping. this fx cdna clone contained 1467 bp of coding sequence, no 5'-untranslated sequence, a short 3'-untranslated sequence of 10 nt and a poly(a) tail at the 3'-end. the fx cdna was inserted into a mammalian expression vector and transfecte ...	1991	1902434
a highly thermostable neutral protease from bacillus caldolyticus: cloning and expression of the gene in bacillus subtilis and characterization of the gene product.	by using a gene library of bacillus caldolyticus constructed in phage lambda embl12 and selecting for proteolytically active phages on plates supplemented with 0.8% skim milk, chromosomal b. caldolyticus dna fragments that specified proteolytic activity were obtained. subcloning of one of these fragments in a protease-deficient bacillus subtilis strain resulted in protease proficiency of the host. the nucleotide sequence of a 2-kb hinfi-mlui fragment contained an open reading frame (orf) that sp ...	1991	1905714
regulation of the chla locus of escherichia coli k12: involvement of molybdenum cofactor.	the chla locus encodes functions required for the biosynthesis of the molybdopterin part of the molybdenum cofactor. mutants, carrying gene fusions at the chla locus, which place beta-galactosidase expression under the control of the chla promoter, have been isolated employing lambda placmu1 as the mutagen. the mutants exhibited beta-galactosidase expression which was greatly enhanced when grown anaerobically. secondary mutations at the chlb, d, e or g loci did not affect the high level of expre ...	1991	1906967
using sodium chloride step gradients to fractionate dna fragments.	a method is described for the separation of dna fragments by ultracentrifugation through a sodium chloride step gradient. the gradients are quickly and easily prepared and require a five- to six-hour centrifugation. fractionated samples of dna may be directly examined by agarose gel electrophoresis, then further analyzed by southern transfer and hybridization. a simple ethanol precipitation followed by several ethanol washes yields fragments that ligate efficiently to vector dna. the method has ...	1991	1907833
the human immunoglobulin kappa locus. characterization of the duplicated o regions.	two large regions of the human immunoglobulin kappa locus, the so-called o regions, have been characterized on cosmid and phage lambda clones. the two regions are very similar but not identical duplicates belonging to the c kappa proximal (p) and the distal (d) copies of the kappa locus. the op and od regions comprise contigs of 90 and 120 kb, respectively, and contain 20 v kappa genes and pseudogenes which have been sequenced. three pairs of v kappa genes were found to be practically identical ...	1991	1907917
in vitro and in vivo analysis of somatic and germline mutability of 2-amino-n6-hydroxyadenine in drosophila melanogaster.	two complementary assays were employed to examine the mutagenicity of 2-amino-n6-hydroxyadenine (aha) in drosophila melanogaster. a lambda phage-based shuttle vector system, utilizing the supf transfer rna gene of escherichia coli, questioned the mutagenicity of aha in established cell cultures derived from somatic tissue while the standard sex-linked recessive lethal assay measured mutational events in vivo. consistent with studies in other systems, aha appears strongly mutagenic when cells are ...	1991	1908052
novel structure of the reca locus of mycobacterium tuberculosis implies processing of the gene product.	a fragment of mycobacterium tuberculosis dna containing reca-like sequences was identified by hybridization with the escherichia coli reca gene and cloned. although no expression was detected from its own promoter in e. coli, expression from a vector promoter partially complemented e. coli reca mutants for recombination, dna repair, and mutagenesis, but not for induction of phage lambda. this clone produced a protein which cross-reacts with antisera raised against the e. coli reca protein and wa ...	1991	1909321
[molecular cloning of alpha-amylase gene from bacillus megaterium and its expression in bacillus subtilis].	using bacteriophage lambda and plasmid pat153 and pnq122 as vectors, alpha-amylase gene from b. megaterium has been cloned into both hosts of e. coli and b. subtilis. expression level of the gene is 250 times higher than b. megaterium when it resides in b. subtilis. the enzyme produced by b. subtilis harboring the hybrid plasmid can digest amylase into maltose and maltotriose at first, then turn them to maltose and glucose, as incubation time extended. it also can digest maltotriose to maltose a ...	1991	1909533
cytoplasmic phosphorylating domain of the mannitol-specific transport protein of the phosphoenolpyruvate-dependent phosphotransferase system in escherichia coli: overexpression, purification, and functional complementation with the mannitol binding domain.	the cytoplasmic c-terminal domain, residues 348-637, and the membrane-bound n-terminal domain, residues 1-347, of eiimtl have been subcloned and expressed in escherichia coli. the n-terminal domain, iicmtl, contains the mannitol binding site, and the c-terminal domain, iibamtl, contains the activity-linked phosphorylation sites, his-554 and cys-384. overexpression of the ba domain was achieved by a translational in-frame fusion of the gene with the cro atg start codon, downstream of the strong p ...	1991	1909895
dna binding properties of the lexa repressor.	the lexa repressor from escherichia coli negatively regulates the transcription of about 20 different genes upon binding with variable affinity to single-, double- or even triple-operators as in the case of the recn gene. binding of lexa to multiple operators is cooperative if the spacing between these operators is favorable. lexa recognizes dna via its amino-terminal domain. the three-dimensional structure of this domain has been determined by nmr measurements. it contains three alpha-helices s ...	1991	1911942
the promoter of the reca gene of escherichia coli.	the growth defect of a lambda phage carrying a reca-lacz fusion was used to select mutations that reduced reca expression. nine single base changes in the reca promoter were isolated that reduced both induced and basal (repressed) levels of expression. deletion analysis of the promoter region and mapping of transcripts indicated that there is one main promoter responsible for both basal and induced expression. some of the mutants displayed a lowered induction ratio, raising the possibility that ...	1991	1911946
regulation of the sos response in bacillus subtilis: evidence for a lexa repressor homolog.	the inducible sos response for dna repair and mutagenesis in the bacterium bacillus subtilis resembles the extensively characterized sos system of escherichia coli. in this report, we demonstrate that the cellular repressor of the e. coli sos system, the lexa protein, is specifically cleaved in b. subtilis following exposure of the cells to dna-damaging treatments that induce the sos response. the in vivo cleavage of lexa is dependent upon the functions of the e. coli reca protein homolog in b. ...	1991	1917874
the murine slp gene. additional evidence that sex-limited protein has no biologic function.	sex-limited protein (slp) is a mouse serum protein of unknown function that has approximately 95% amino acid sequence identity with murine complement component c4 but is inactive in the complement pathway. the gene for slp lies in the s region of the murine h-2 complex adjacent to the gene cyp21 that encodes the cytochrome p-450 enzyme steroid 21-hydroxylase. we report the sequence of a 26,307 bp long segment of the mouse genome that includes both the slp and cyp21 genes. the sequence reported w ...	1991	1918990
regulation of the immune response to peptide antigens: differential induction of immediate-type hypersensitivity and t cell proliferation due to changes in either peptide structure or major histocompatibility complex haplotype.	the immunodominant cd4 t cell epitope of the bacteriophage lambda ci repressor protein in several inbred mouse strains can be represented by a peptide encompassing amino acids 12-26. here, we show that this peptide, and a variety of its sequence variants, can induce immediate-type hypersensitivity in mice. 12-26 variants that differ by as little as single amino acid residues deviate greatly in their ability to induce hypersensitivity. further, differences in major histocompatibility complex clas ...	1991	1919438
escherichia coli xerc recombinase is required for chromosomal segregation at cell division.	xerc is a site-specific recombinase of the bacteriophage lambda integrase family that is encoded by xerc at 3700 kbp on the genetic map of escherichia coli. the protein was originally identified through its role in converting multimers of plasmid cole1 to monomers; only monomers are stably inherited. here we demonstrate that xerc also has a role in the segregation of replicated chromosomes at cell division. xerc mutants form filaments with aberrant nucleotides that appear unable to partition cor ...	1991	1931824
molecular cloning and expression of rat placental lactogen-iv, a variant of rpl-i present in late pregnant rat placenta.	late pregnant rat placenta was found to contain a messenger rna (mrna) that encodes an additional member of the prolactin-growth hormone family. this polypeptide was detected by hybridization of complementary dna (cdna) for rpl-i to a 1 kilobase mrna transcript in late pregnant rat placenta. a cdna clone for the new polypeptide was isolated from a phage lambda-gt10 library containing cdna synthesized from day 18 rat placental mrna. sequencing of the day 18 cdna clone revealed that it was most cl ...	1991	1935804
determination of the cdna sequence for the human mitochondrial 75-kda fe-s protein of nadh-coenzyme q reductase.	a human-hepatoma cdna lambda gt11 expression library was probed with an antibody to holoenzyme complex i (nadh-coq reductase) of the respiratory chain. one of the 30 antibody positive clones was purified to homogeneity, amplified by the polymerase chain reaction (pcr), subcloned and sequenced. it proved to be highly similar to the cdna sequence for the bovine 75-kda fe--s protein. using the sequence obtained from this library, both sense and antisense oligonucleotides were constructed and used t ...	1991	1935949
acidic pentapeptide phosphorylated in vitro by calf thymus protein kinase nii binds to dna in the presence of mg2+ cations.	the pentapeptide pyroglu-ala-glu-ser-asn has been synthetized and phosphorylated in vitro at level of serine by protein kinase nii isolated from calf thymus chromatin. it is noteworthy that the calf thymus kinase nii shows a remarkable affinity for this peptide. the [32p]peptide is able to bind to several dnas in the presence of mg2+ (lambda phage, calf thymus, pbr540 plasmid). this binding appears not specific with regard to the type of dna and its base sequence. these data support the hypothes ...	1991	1936253
the isolation and characterization of a calmodulin-encoding gene (cmd1) from the dimorphic fungus candida albicans.	candida albicans is a dimorphic, opportunistic pathogen of humans, and calcium and calmodulin have been implicated in its morphogenic transition. the c. albicans calmodulin-encoding gene, cmd1, was isolated from cdna and genomic phage lambda libraries using the saccharomyces cerevisiae cmd gene as a hybridization probe. southern-blot hybridization analysis of genomic dna suggests the existence of only one type of calmodulin gene in c. albicans. comparison of cdna and genomic sequences identified ...	1991	1937040
murein-metabolizing enzymes from escherichia coli: sequence analysis and controlled overexpression of the slt gene, which encodes the soluble lytic transglycosylase.	the complete nucleotide sequence of the slt gene encoding the soluble lytic transglycosylase (slt; ec 3.2.1.-) from escherichia coli has been determined. the largest open reading frame identified on a 2.5-kb pvuii-sali fragment indicates that the enzyme is translated as a preprotein of either 654 or 645 amino acids, depending on which of two potential start codons is used. the two possible translation products differ only in the lengths of their predicted signal peptides, 36 or 27 amino acids, r ...	1991	1938883
different interactions of cro repressor dimer with the left and right halves of or3 operator dna.	lambda cro repressor protein is titrated with two half-operator dna duplexes comprising the right and left halves of the major binding site on phage lambda dna, the or3 operator. the comparison of binding strengths and the conformation of cro repressor in the two protein-dna complexes shows that base pair differences between the two halves of the or3 operator affect the binding of cro repressor protein. some 1h nmr resonances are assigned for both protein and dna in the cro-operator dna complexe ...	1991	1939232
high somatic mutation frequencies in a lacz transgene integrated on the mouse x-chromosome.	to study spontaneous and induced mutagenesis in vivo we recently constructed a series of transgenic mice harboring different numbers of bacteriophage lambda shuttle vectors, provided with a lacz mutational target gene, integrated in their genome. the transgenic mice enabled analysis of spontaneous and induced mutation frequencies in postmitotic tissues like liver and brain. the obtained data indicated spontaneous mutation frequencies in the order of 10(-5)-10(-6). here we report a 25-100 times h ...	1991	1944355
cloning, sequencing and overexpression of the gene for prokaryotic factor ef-p involved in peptide bond synthesis.	a soluble protein ef-p (elongation factor p) from escherichia coli has been purified and shown to stimulate efficient translation and peptide-bond synthesis on native or reconstituted 70s ribosomes in vitro. based on the partial amino acid sequence of ef-p, 18- and 24-nucleotide dna probes were synthesized and used to screen lambda phage clones from the kohara gene bank. the entire ef-p gene was detected on lambda clone #650 which contains sequences from the 94 minute region of the e.coli genome ...	1991	1956781
tritylase antibodies.	we have used a tris(4-methoxyphenyl)-phosphonium compound as a hapten to elicit catalytic antibodies that selectively remove trityl protecting groups at neutral ph. one antibody, 37c4, was characterized kinetically with a number of trityl substrates. the rate enhancement was consistently near 200; the km was approximately 30 microm for the methoxytrityl substrates. compounds with no methoxy substituents on the trityl group were not hydrolysed by the antibody. no decrease in the rate of reaction ...	1991	1959450
selective cleavage of human dna: reca-assisted restriction endonuclease (rare) cleavage.	current methods for sequence-specific cleavage of large segments of dna are severely limited because of the paucity of possible cleavage sites. a method is described whereby any eco ri site can be targeted for specific cleavage. the technique is based on the ability of reca protein from escherichia coli to pair an oligonucleotide to its homologous sequence in duplex dna and to form a three-stranded complex. this complex is protected from eco ri methylase; after methylation and reca protein remov ...	1991	1962209
a cdna encoding 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase from solanum tuberosum l.	a cdna encoding potato (solanum tuberosum l.) 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase, the first enzyme of the shikimate pathway, was cloned into phage lambda gt11. the clone represents the first cdna for this enzyme from any eukaryotic source. the nucleotide sequence of the cdna was determined, and its identity was confirmed through partial amino acid sequence analysis of the encoded enzyme. the cdna contains a 1527-base pair open reading frame that encodes a polypeptide with a cal ...	1990	1967256
isolation of anonymous, polymorphic dna fragments from human chromosome 22q12-qter.	a series of 195 random chromosome 22-specific probes, equivalent to approximately 1% of the size of this chromosome, have been isolated from a chromosome 22-specific bacteriophage lambda genomic library. these probes were mapped to four different regions of chromosome 22 on a panel of five somatic cell hybrids. restriction fragment length polymorphisms were detected by 28 of the probes mapping to 22q12-qter. evolutionarily conserved sequences in human, mouse, and chinese hamster dna were detecte ...	1990	1968030
isolation and characterization of the rat glutamine synthetase-encoding gene.	from a rat genomic library in phage lambda charon4a, a complete glutamine synthetase-encoding gene was isolated. the gene is 9.5-10 kb long, consists of seven exons, and codes for two mrna species of 1375 nucleotides (nt) and 2787 nt, respectively. for both mrnas, full-length cdnas containing a short poly(a) tract were identified. the sequences of the entire mrna and of the exon-intron transitions were determined. the smaller mrna is identical to the 5' 1375 nt of the long mrna and contains the ...	1990	1970548
characterization of the pilin gene of moraxella bovis dalton 2d and expression of pili from m. bovis in pseudomonas aeruginosa.	the pilin gene of moraxella bovis dalton 2d was isolated by cloning in pseudomonas aeruginosa. the nucleotide sequence of this gene encodes a prepilin of 156 amino acid residues. when high levels of pilin were expressed from the gene in p. aeruginosa, by using the pl promoter of bacteriophage lambda inserted upstream of the coding sequence, pili which were indistinguishable from pili of m. bovis were produced.	1990	1971258
structure and polymorphic map of human lipoprotein lipase gene.	lipoprotein lipase (lpl) catalyzes the key step for the removal of triacylglycerol-rich lipoproteins from the circulation. in this paper, we report the cloning and structure of the normal human lpl gene, which was isolated in three overlapping lambda phage clones that span about 35 kilo bases (kb) of the genetic locus. the peptide coding region of the gene is approx. 23 kb in length and contains nine exons with intron sizes ranging from 0.7 to 8.7 kb. the entire 3' untranslated region is in the ...	1990	1972631
the murine genes hox-5.1 and hox-4.1 belong to the same hox complex on chromosome 2.	two different loci of antennapedia-related homeobox-containing genes have been shown to map to mouse chromosome 2: the hox-5 complex and the hox-4.1 gene. these independently derived loci are likely to be parts of a single gene complex, although their close linkage has not yet been demonstrated. since cosmid walks to extend the hox-5 cluster and to potentially link the two loci were unsuccessful, we have used large restriction fragments separated by pulsed-field gel electrophoresis to demonstrat ...	1990	1973141
a computer program to assist in the choice of restriction endonucleases for use in dna analyses.	type ii restriction endonucleases cleave double stranded dna molecules at sites characterized by one or more sets of nucleotide pairs sequences. these digestions are essential in such procedures as dna cloning, dna sequencing and restriction fragment length polymorphism (rflp) analyses. a large number of enzymes with different sequence specificities are available. to date, most choices of restriction endonucleases have been made by trial and error. a computer program, redi, has been developed th ...	1990	1975563
transposon-induced non-motile mutants of vibrio cholerae.	non-motile mutants of vibrio cholerae were isolated after transposon insertion mutagenesis with either tn5 on a plasmid or tn10ptac mini-kan in bacteriophage lambda. the physical location and number of transposon insertions was determined. eighteen tn5 insertion mutants and 11 tn10ptac mini-kan insertion mutants had single unique insertion sites. the 18 tn5 insertions were contained within six different ecori fragments and the 11 tn10ptac mini-kan insertions were contained within eight different ...	1990	1975833
mapping of insertion element is30 in the escherichia coli k12 chromosome.	we identified seven phage clones containing the insertion element is30 in a lambda phage library mini-set, which includes 476 clones carrying chromosomal segments that cover almost the entire chromosome of escherichia coli k12 w3110 (kohara et al. 1987). we could assign locations and orientations to four copies of is30 (named is30a to is30d) on the w3110 chromosome by restriction analysis of phage dnas containing them. these is30s were present at the same locations in chromosomes of both w3110 a ...	1990	1980336
comparison of the action of ionizing radiation and uv-light on lambda phage. influence on phage adsorption, dna injection, replication, and dna repair.	the influence of gamma radiation, x-rays, uv-light at 254 nm and 365 nm, the latter combined with furocoumarin sensitizers has been studied on plaque forming ability, phage adsorption, dna injection, and replication processes. uv-light (365 nm) plus furocoumarin treatment of phage particles gave rise to two types of dna crosslinks. type i crosslink corresponded to furocoumarin mediated covalent linkage between adjacent sites in opposite strands of the double helix. crosslink type ii (hairpin cro ...	1990	1981452
evolutionary aspects, structure, and expression of the rat interleukin 4 gene.	the rat interleukin 4 (il 4) gene has been isolated from a genomic lambda phage library by cross-hybridization to the mouse il 4 cdna. like the mouse and human counterparts, it exists as a single copy gene in the genome and consists of four exons. the overall structure of the il 4 locus seems highly conserved. this is indicated by the low degree of restriction fragment length polymorphism in a number of laboratory and wild mice and by the conservation of the intron size between human, rat, and m ...	1990	1983334
fibrinogen is not synthesized by human megakaryocytes.	the origin of platelet fibrinogen is controversial. it may arise from two sources: (a) exogenously by endocytosis of plasma fibrinogen, or (b) endogenously by synthesis. we explored the second possibility because we previously demonstrated that the first mechanism does occur. fibrinogen synthesis by human megakaryocytes (mk) was investigated by in situ hybridization and the polymerase chain reaction (pcr) applied to mrna. mk differentiating from marrow cfu-mk were cultured in suspension. in situ ...	1991	1985697
structure of the human laminin b2 chain gene reveals extensive divergence from the laminin b1 chain gene.	the exon-intron structure of the human laminin b2 chain gene was elucidated from genomic lambda phage clones spanning 2 kilobase pairs (kb) of the 5'-flanking region, 58 kb of the structural gene and 10 kb of the 3'-flanking region. the entire gene was shown to contain 28 exons. the promoter region has no tata or caat boxes whereas it contains five gc boxes and three ap-2-like binding sites. comparison with the promoter region of the mouse gene revealed six highly conserved sequences of 14 to 42 ...	1991	1985895
role of a disulfide bond in the thermal stability of the lamb protein trimer in escherichia coli outer membrane.	in order to understand the unusual heat resistance of lamb protein (the outer membrane component of the maltose transport system in escherichia coli and its receptor for bacteriophage lambda), we investigated the role of its 2 cysteinyl residues. our studies show that cys22 and cys38 form an intrasubunit disulfide bond which contributes to the heat stability of the lamb protein trimer. physical evidence for the disulfide was obtained by using site-directed mutagenesis to convert asn36 to met, wh ...	1991	1988451
isolation and sequencing of cdna clones encoding the dictyostelium discoideum 30,000-dalton actin-bundling protein.	the dictyostelium 30,000-dalton protein is a calcium-regulated actin filament-bundling protein which has been suggested to contribute to the structure and reorganization of filopodia and pseudopodia accompanying cell movements. cdnas encoding this protein were isolated using antibody and oligonucleotide probes to screen cdna libraries in phage lambda. the sequence of the cdna predicts a protein of 295 amino acids with a molecular weight of 33,355. the sequence reveals two ef-hand calcium-binding ...	1991	1993662
cloning and overexpression of the gene encoding bacteriophage t5 dna polymerase.	t5 dna polymerase (t5pol), an essential enzyme for bacteriophage t5 dna replication, is unusual because of its high processivity and strand-displacing ability. these two properties in a single polypeptide make t5pol an ideal candidate for structural and functional analysis. therefore, the structural gene encoding the dna polymerase of bacteriophage t5 (t5pol) has been cloned and overexpressed in escherichia coli. elimination of sequences upstream from the 5' end of the t5pol by exonuclease iii d ...	1991	1995424
a homologue of the bacterial heat-shock gene dnaj that alters protein sorting in yeast.	heat-shock proteins have been implicated in assembly of protein complexes, correct protein folding and uptake of proteins into organelles. in escherichia coli, the heat-shock protein dnaj and the hsp70 homologue, dnak, act together to disassemble a protein complex involved in bacteriophage lambda replication. we report the identification of scj1, a gene in the yeast saccharomyces cerevisiae that encodes a homologue of the bacterial dnaj protein. scj1 was identified by a genetic screen in which i ...	1991	2000136
characterization of the 3' half of the human type iv collagen alpha 5 gene that is affected in the alport syndrome.	we have determined the exon-intron structure of the 3' half of the gene for the human type iv collagen alpha 5 chain that is affected in x-chromosome-linked alport syndrome. six overlapping lambda phage genomic clones containing exons 1-14 (as counted from the 3' end) and two additional overlapping genomic clones containing exons 16-19 spanned a total of 60 kb, 9.5 kb of which were the 3' flanking region. the exon-intron structure was elucidated by restriction enzyme mapping, nucleotide sequenci ...	1991	2004755
cloning and characterization of a human c-myc promoter-binding protein.	a human cdna clone encoding a c-myc promoter-binding protein was detected by screening a hela cell lambda phage expression cdna library. the library was screened by using an xhoi-naei human c-myc p2 promoter fragment as a probe. the recombinant phage encoded a fusion protein, myc-binding protein 1 (mbp-1), which had an apparent molecular size of 40 kda. a corresponding protein with a molecular size of 35 kda was present in a hela cell extract. sequence analysis of the cloned gene reveals an open ...	1991	2005901
function of dnaj and dnak as chaperones in origin-specific dna binding by repa.	heat-shock proteins are normal constituents of cells whose synthesis is increased on exposure to various forms of stress. they are interesting because of their ubiquity and high conservation during evolution. two families of heat-shock proteins, hsp60s and hsp70s, have been implicated in accelerating protein folding and oligomerization and also in maintaining proteins in an unfolded state, thus facilitating membrane transport. the escherichia coli hsp70 analogue, dnak, and two other heat-shock p ...	1991	2005967
high-level expression of a semisynthetic dam gene in escherichia coli.	we constructed a semisynthetic gene encoding a dna-adenine-methyltransferase (dam) that codes for the same amino acid sequence as the wild type (wt) escherichia coli dam gene. since for unknown reasons the entire wt sequence, from the start codon to the end of the gene, could not be cloned, a gene was constructed consisting of a chemically synthesized 5' portion and a 3' portion from the e. coli chromosome. introduction of this semisynthetic gene into a suitable vector allows overproduction of e ...	1991	2013413
structural and functional comparison between the stability systems pard of plasmid r1 and ccd of plasmid f.	the stability determined by the systems pard of plasmid r1 and ccd of plasmid f is due to the concerted action of two proteins, a cytotoxin and an antagonist of this function. in this paper we report that ccda and kis proteins, the antagonists of the ccd and pard systems respectively, share significant sequence homologies at both ends. in kis, these regions seem to correspond to two different domains. despite the structural similarities, kis and ccda are not interchangeable. in addition we have ...	1991	2017133
cloning and sequencing of an escherichia coli k12 gene which encodes a polypeptide having similarity to the human ferritin h subunit.	using lambda phage clones containing segments of the escherichia coli k12 chromosome as hybridization probes, we found one gene at 42 min on the e. coli chromosome map, the expression of which was affected by rnase iii. the sequence of the dna fragment containing this gene (gen-165) revealed the presence of an open reading frame encoding a polypeptide of 165 amino acid residues. the amino acid sequence deduced from the nucleotide sequence exhibited a remarkable similarity to that of the human fe ...	1991	2017145
reca-independent resistance to irradiation with u.v. light in acid-habituated escherichia coli.	growth of escherichia coli 1829 colv, i-k94 at ph 5.0 led to an increase in u.v. resistance compared with cells grown at ph 7.0. this was due to a phenotypic change, since organisms grown at ph 7.0 showed increased resistance after only 2.5-5.0 min incubation at the mildly acid ph. other e. coli k12 derivatives became more u.v.-resistant at ph 5.0 including uvra, reca and pola1 mutants. organisms grown at ph 5.0 also showed increased weigle reactivation of u.v.-irradiated lambda phage and this a ...	1991	2019551
dual start motif in two lambdoid s genes unrelated to lambda s.	the lysis gene region of phage 21 contains three overlapping reading frames, designated s21, r21, and rz21 on the basis of the analogy with the srrz gene cluster of phage lambda. the 71-codon s21 gene complements lambda sam7 for lysis function but shows no detectable homology with s lambda in the amino acid or nucleotide sequence. a highly related dna sequence from the bacteriophage pa-2 was found by computer search of the genbank data base. correction of this sequence by insertion of a single b ...	1991	2019562
the u1 snrna gene repeat from the sea urchin (strongylocentrotus purpuratus): the 70 kilobase tandem repeat ends directly 3' to a u1 gene.	lambda phage clones containing multiple copies of the 1.1 kb tandemly repeated unit of the sea urchin (s. purpuratus) u1 rna genes were isolated from a gene library. the 1.1 kb repeat unit encodes a single copy of the predominant u1 rna expressed in oocytes and embryos prior to the blastula stage. the tandem repeat unit is about 80 kb in size and is probably present one time per haploid genome as judged by pulsed-field electrophoresis of sperm dna digested with restriction enzymes which do not c ...	1991	2020546
design of the helix-turn-helix motif: nonlocal effects of quaternary structure in dna recognition investigated by laser raman spectroscopy.	the operator-binding domain of phage lambda repressor provides a model for dna recognition by the helix-turn-helix (hth) motif. in the wild-type protein, dimerization is mediated by hydrophobic packing (of the dyad-related helix 5), which serves as an indirect determinant of operator affinity. the mutant repressor, tyr88----cys, forms an intersubunit disulfide linkage and exhibits enhancement of both structural stability and operator affinity. yet the dimer-specific operator affinity of the muta ...	1991	2021630
molecular cloning of dna from a sorted human minichromosome.	a human supernumerary minichromosome (mc), found in a newborn baby and sorted on a fluorescence-activated cell sorter (facs-440) has been previously described [ferretti et al., cytotechnology 1 (1987) 7-12]. we report here on the construction of a library of ecori fragments in the phage lambda gtwes.lambda b', starting from 7.5 ng of mc dna, and describe the isolation of single-copy dna clones from the library in a two-step procedure. we employed in situ hybridization to unambiguously select the ...	1991	2022335
molecular cloning and characterization of the reca gene of rhizobium phaseoli and construction of reca mutants.	the rhizobium phaseoli reca gene has been cloned by interspecific complementation of the fec phenotype of bacteriophage lambda. the cloned gene restored the recombination proficiency and conferred resistance to dna-damaging agents (methyl methanesulfonate and nitrofurantoin) to an escherichia coli reca mutant. the direction of transcription and the localization of the reca gene were determined by mutagenesis with phage mudiipr13 and heterologous hybridization with an e. coli reca probe. an r. ph ...	1991	2022610
subcloning, nucleotide sequence, and expression of trkg, a gene that encodes an integral membrane protein involved in potassium uptake via the trk system of escherichia coli.	the trkg gene encodes a component of the k+ uptake system trk and is located at 30.5 min inside the lambdoid prophage region rac of the escherichia coli chromosome. trkg was subcloned, its nucleotide sequence was determined, and its product was identified in a minicell system. the open reading frame of 1,455 bp encodes a hydrophobic membrane protein with a calculated molecular weight of 53,493 that is predicted to contain up to 12 transmembrane helices. the trkg gene product behaved as a hydroph ...	1991	2022616
a 14-kda schistosoma mansoni polypeptide is homologous to a gene family of fatty acid binding proteins.	the complete nucleotide sequence encoding a schistosoma mansoni protein termed sm14 was determined from cdna clones propagated in bacteriophage lambda gt11 in escherichia coli. the 14.8-kda protein bears significant homologies with a family of related polypeptides which bind hydrophobic ligands. members of this group of cytosolic proteins were originally identified based on their affinity for long chain fatty acids. the purified recombinant protein exhibited an affinity to fatty acids, in contra ...	1991	2022660
combinatorial mutagenesis of the reactive site region in plasminogen activator inhibitor i.	plasminogen activator inhibitor (pai-i) rapidly inactivates tissue plasminogen activator (t-pa) and urokinase (uk) with nearly identical association rate constants. the contributions of ser344, ala345, and arg346 (p3, p2, and p1 residues, respectively) in pai-i to inhibition of uk and t-pa were evaluated using combinatorial mutagenesis of the human pai-i cdna. a bacteriophage lambda expression library potentially encoding the 8000 unique pai-i species were screened for inhibitory activity agains ...	1991	2022663
construction and characterization of a single-chain catalytic antibody.	the antigen-binding (fab) fragment of the catalytic monoclonal antibody npn43c9 has recently been cloned by using bacteriophage lambda. by inserting the variable regions of this fab coding sequence into a (nh2)-vl-linker-vh-(cooh) construct (where vl and vh represent the heavy and light chain variable regions), we have assembled a recombinant gene encoding a catalytic single-chain antigen-binding protein. this protein has been expressed in escherichia coli and exhibits the same catalytic paramet ...	1991	2023948
tolbutamide and phenytoin hydroxylations by cdna-expressed human liver cytochrome p4502c9.	a human cytochrome p4502c9 cdna clone has been isolated from a human liver bacteriophage lambda gt11 library using oligonucleotide probes. expression of the 1762 base pair cdna in cos cells demonstrated that the encoded enzyme has a molecular mass of 55 kda as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. the expressed enzyme catalysed the methylhydroxylation of tolbutamide with an apparent km of 131.7 microm, similar to that observed in human liver microsomes. p4502c ...	1991	2025243
polymerase chain reaction for detection of mycoplasma gallisepticum.	a species-specific 760-base pair (bp) bamhi to ecori dna fragment (fmg-2) was isolated from a mycoplasma gallisepticum (mg) genomic library constructed in plasmid puc8. based on the dna sequence data of fmg-2, a pair of 25 base primers, designated amplification (amp) left (l) and right (r) primers, was synthesized. when used in the polymerase chain reaction (pcr), the amp l and r primers directed amplification of dna of 16 mg strains yielding an expected 732-bp product, but did not amplify dna o ...	1991	2029263
[inhibitory effects of chinese medicines on sos responses in e. coli and their mechanism].	sixty kinds of commonly used chinese medicines have been examined for their ability to depress the release of lambda phage from lysogenic strain in the inductest. 11 chinese medicines showed an inhibitory effects. among them, codonopsis radix, polygonatum radix and fractus lycium were strong depressors. they also showed an inhibitory effect on sos response in sos chromotest with a dose-effect response. these medicines were also found to decrease the frequency of gene conversion in s. cerevisiae ...	1991	2029430
construction of an ordered clone bank and systematic analysis of the whole transcripts of chromosome vi of saccharomyces cerevisiae.	by comparing sequences of restriction enzyme cleavage sites and their distance data, we sorted 384 lambda phage clones containing segments of chromosome vi of s. cerevisiae and constructed an ordered clone bank for this chromosome. the physical length of this bank is 269.7 kb. the bank contains the entire chromosome including the left telomere, but it is not certain whether it contains the right telomere as well. to estimate the number of genes present on this chromosome, we performed a series o ...	1991	2030939
a method of identifying and isolating a unique member of a multigene family: application to a trypanosome surface antigen gene.	a chimeric oligonucleotide was constructed using dna sequences from two distal regions of a cdna which encodes a major surface antigen (tsa-1) of trypanosoma cruzi. conditions were found that allowed the chimeric oligonucleotide to hybridize only to a 5.4 kb ecori fragment in a southern blot of total genomic dna. the 5.4 kb ecori genomic dna fragment has previously been shown to be located at a telomeric site, thus the studies described here directly demonstrate that the tsa-1 gene is telomeric ...	1991	2030963