Publications

resolution of synthetic att-site holliday structures by the integrase protein of bacteriophage lambda.	site-specific recombination of the bacteriophage lambda genome into and out of the host bacterial genome is postulated to involve the formation of holliday structure intermediates by reciprocal single-strand exchanges. synthetic analogues of the predicted recombination intermediates are resolved in vitro by the protein product of the lambda int gene. some of the structural features and reaction conditions for this genetic recombination can now be defined.	1984	6092961
controlled synthesis of the coat protein of satellite tobacco necrosis virus in escherichia coli.	chimeric plasmids were constructed such that the cloned complete satellite tobacco necrosis virus (stnv) rna information came under transcriptional control of the leftward promoter (pl) of bacteriophage lambda. the promoter is fully repressed at low temperatures (28 degrees) by the thermolabile repressor product of the lambdaci857 gene, present in the bacterium on a deficient prophage or as part of another plasmid. synthesis of the stnv coat protein in escherichia coli could be initiated by heat ...	1984	18639819
assembly pathway of newly synthesized lamb protein an outer membrane protein of escherichia coli k-12.	the assembly of newly induced lamb protein (phage lambda receptor) was investigated in an operon fusion strain of escherichia coli, in which the lamb gene is expressed under lac promoter control. the induction kinetics both for total cellular and for cell surface-exposed lamb protein were studied by immunochemical detection methods, using two distinct antisera directed against detergent-solubilized lamb trimers and completely denatured lamb monomers, respectively. anti-trimer antibodies recogniz ...	1984	6204059
identification of transfer rna suppressors in escherichia coli. iv. amber suppressor su+6 a double mutant of a new species of leucine trna.	an escherichia coli dna fragment containing an su+6 amber suppressor gene (supp) was cloned into a lambda gt lambda ch vector by the shotgun method, selecting a su+6 transducing phage lambda psu+6. through prophage integration followed by induction occurring at the transducing region of the lambda psu+6 in su- e. coli, a counterpart transducing phage carrying the wild-type allele (su degrees 6) was isolated (lambda psu degrees 6). the fingerprint of a trna encoded by lambda psu degrees 6 was ide ...	1984	6207302
cloning, nucleotide sequence and high level expression of the gene coding for the connector protein of bacillus subtilis phage phi 29.	the phi 29 dna restriction fragment hindiii-d, shown to contain gene 10 coding for the connector protein, has been cloned in plasmid pplc28 under the control of the pl promoter of phage lambda. after heat induction to inactivate the lambda repressor, a protein with the electrophoretic mobility of the connector protein p10 was synthesized, accounting for about 30% of the total escherichia coli protein after 3 h of induction. the 2205 nucleotide-long sequence of the cloned hindiii-d fragment has b ...	1984	6096227
lack of induction of non-targeted mutations in intact bacteriophage by uvb (313 nm), uva (334 nm, 365 nm) and visible (405 nm) irradiation of host cells.	mutation to virulence has been measured in intact bacteriophage lambda 15 infected into host cells pre-treated with uvc (254 nm), uvb (313 nm), uva (334 nm, 365 nm) or visible (405 nm) radiations. we have confirmed that uvc radiation leads to a large enhancement (maximum enhancement factor of 140 in wild-type) of the background spontaneous mutation frequency (non-targeted mutagenesis) and have further shown that this is at least partially dependent on excision repair (maximum enhancement factor ...	1984	6229698
replication defective rp4 plasmids recovered after chromosomal integration.	phh6000 is a composite replicon made by the in vitro ligation of the incp plasmid rp4 to a fragment of bacteriophage lambda capable of autonomous replication. derivatives were selected in which it had integrated into the escherichia coli chromosome by homologous recombination with the resident lambda prophage, and plasmids were subsequently regenerated from the integrated molecules. although of the same molecular size as phh6000, all had altered properties: those recovered from the chromosome of ...	1984	6231650
facile and gentle method for quantitative lysis of escherichia coli and salmonella typhimurium.	garrett et al. (mol. gen. genet. 182:326-331, 1981) constructed strains of escherichia coli harboring derivatives of plasmid pbr322 that carry the lysis genes (s, r, and rz) of phage lambda. the plasmid construction placed the genes under control of the lactose operon operator-promotor (and thus of lac repressor). induction of e. coli strains carrying these plasmids resulted in rapid lysis of the culture unless the s gene was defective, in which case the cells grew normally. a freeze-thaw treatm ...	1984	6232260
an integration-proficient int mutant of bacteriophage lambda.	we have isolated and characterized a novel int mutant of phage lambda. this mutant promotes efficient recombination between the phage and bacterial attachment sites, but, unlike wild type, does not promote efficient recombination of any other pair of attachment sites tested in most conditions. in particular, recombination between two phage or two prophage attachment sites is poor relative to the wild type frequency. we attribute this unusual phenotype to differences in the distribution of int pr ...	1984	6238223
genetic analysis of clear-plaque mutations induced in bacteriophage lambda by 9-aminoacridine.	clear-plaque mutations were induced in the ci and cii genes of lambda by treating lysogenic cells with 9-aminoacridine (9aa). mapping of the mutations revealed that there were two hot spots for 9aa mutagenesis in ci, and one strong hot spot in cii. the hot spots in ci mapped close to 1 of the 3 runs of 4 g/c base-pairs and near the only run of 5 g/cs, respectively, in this gene. of 36 ci mutations tested, at most one mapped near a run of 6 a/t base-pairs. by analogy, the sequence responsible for ...	1984	6239977
mutations in the dna gyrb gene that are temperature sensitive for lambda site-specific recombination, mu growth, and plasmid maintenance.	we report the isolation of two mutations in the gyrb gene of escherichia coli k12 obtained from an initial selection for resistance to coumermycin a1 and a subsequent screening for bacteria that fail to support site-specific recombination of phage lambda, i.e., him-. these two mutations have a temperature-sensitive him- phenotype, supporting site-specific recombination efficiently at low temperature, but inefficiently at high temperatures. like other him mutants, the gyrb-him mutants fail to pla ...	1984	6319362
cloning of the dnab gene of escherichia coli: the dnab gene of gropb534 and gropb612 and the replication of phage lambda.	fragments of the e. coli chromosome that carry the dnab gropb534 or gropb612 alleles have been cloned into a cosmid vector. the resulting recombinant plasmids contained the genes uvra, grop (b534 or b612), and lexa. further subcloning into high copy number plasmids, during which the uvra and lexa genes were removed successively, yielded a gropb534 and gropb612 dna fragment of about 2.4 kb each. both fragments contained an overlapping 1.8 kb segment of dna in which the sites of all restriction en ...	1984	6319960
mrna processing in escherichia coli: an activity encoded by the host processes bacteriophage f1 mrnas.	to examine the regions of the male-specific filamentous bacteriophage f1 genome that include signals for mrna processing, the 5' endpoints of the major in vivo phage mrnas have been located in the f1 dna sequence by s1 nuclease mapping. the 5' ends of the purified mrnas and additional phage-specific rnas transiently visible early after infection occur in clusters of t-rich residues within genes that code for three phage proteins. when a 270-nucleotide region encompassing the 5' endpoints of thre ...	1984	6322124
[c1 and cro repressors of lambda phages. i. construction of vectors for expression of cro repressor of bacteriophage lambda imm434].	eight derivatives of recombinant plasmid pbrcro434, that consists of pbr322 and fragment of immunity region of phage lambda imm434 have been constructed and characterised. these derivatives contain the deletions in the region adjacent to or3 operator and in the structural gene of cro-repressor of lambda imm434. the deletions have been produced by the treatment of pbrcro434 with exonuclease iii of escherichia coli and s1 nuclease of aspergillus orizae and precisely mapped. the unique ecori-restri ...	1984	6323976
cloning and manipulation of the escherichia coli cyclopropane fatty acid synthase gene: physiological aspects of enzyme overproduction.	like many other eubacteria, cultures of escherichia coli accumulate cyclopropane fatty acids (cfas) at a well-defined stage of growth, due to the action of the cytoplasmic enzyme cfa synthase. we report the isolation of the putative structural gene, cfa, for this enzyme on an e. coli-cole1 chimeric plasmid by the use of an autoradiographic colony screening technique. when introduced into a variety of e. coli strains, this plasmid, plc18-11, induced corresponding increases in cfa content and cfa ...	1984	6325391
lambda placmu: a transposable derivative of bacteriophage lambda for creating lacz protein fusions in a single step.	we isolated a plaque-forming derivative of phage lambda, lambda placmu1 , that contains sequences from bacteriophage mu enabling it to integrate into the escherichia coli chromosome by means of the mu transposition system. the mu dna carried by this phage includes both attachment sites as well as the ci, ner (cii), and a genes. lambda placmu1 also contains the lacz gene, deleted for its transcription and translation initiation signals, and the lacy gene of e. coli, positioned next to the termina ...	1984	6327627
identification of the phom gene product and its regulation in escherichia coli k-12.	plasmids containing the chromosome region of escherichia coli encoding phom, whose product is a positive regulator of alkaline phosphatase expression, were isolated from the clarke and carbon plasmid bank. a 9.9-kilobase ecori fragment of plasmid plc17-39 (subcloned into pbr322) was able to complement both phom and thrb mutations. restriction endonuclease analysis and in vitro mutagenesis of the hybird plasmids enabled the localization of the phom gene locus to 3 kilobases of the cloned chromoso ...	1984	6330029
[mutagenic effects of gamma-rays on plasmid dna in escherichia coli].	a purified and dried dna of plasmid pko482 (galk+) is 10 times more resistant to the inactivating action of 60co-gamma-rays than that of lambda phage. gamma-irradiation of the plasmid dna induces forward mutations of galk, the frequency of which increases linearly with the dose. the efficiency of the mutagenic action of gamma-rays on the plasmid galk locus is 10(-12) per 1 rad and per 1 base pair. the mutagenic effect of gamma-radiation but slightly depends upon bacterial reca+ gene and upon the ...	1984	6390494
mapping of the glucose dehydrogenase gene in bacillus subtilis.	a 4.0-kilobase dna fragment containing the developmentally regulated gene for glucose dehydrogenase (gdh) from bacillus subtilis was incorporated into the plasmid pgx345, which contains a marker conferring chloramphenicol resistance (cat). the resistance marker of the resulting integration vector was used to map the gdh gene on the b. subtilis chromosome. using pbs1 transduction, the gene order was determined to be aroi cat (gdh) mtlb dal. the cat (gdh) marker was also cotransformable with mtlb. ...	1984	6438057
construction of a plasmid that overproduces the large proteolytic fragment (klenow fragment) of dna polymerase i of escherichia coli.	using currently available gene fusion techniques, we have constructed plasmids that direct the overproduction of the carboxyl-terminal two-thirds of dna polymerase i, corresponding to the proteolytically derived "klenow fragment." we have obtained overproduction amounting to several percent of the cellular protein using constructs in which expression is directed either from the lac promoter or from the leftward promoter of phage lambda. the polymerase fragment has been purified to homogeneity fr ...	1983	6340110
stabilization of proteins by a bacteriophage t4 gene cloned in escherichia coli.	the cloned bacteriophage t4 pin gene functions to stabilize several different kinds of proteins in escherichia coli bacteria. incomplete proteins such as puromycyl polypeptides, abnormal but complete proteins such as the lambda phage tso protein, and labile eukaryotic proteins encoded by genes cloned in e. coli such as mature human fibroblast interferon are stabilized in cells in which the t4 pin gene is expressed. the cloned t4 pin gene does not seem to affect the turnover of normal e. coli pro ...	1983	6340113
expression of cloned mitochondrial dna from the petite negative yeast schizosaccharomyces pombe in e. coli minicells.	the minicell producing escherichia coli strain d24 (lysogenic for phage lambda ci857) was transformed with the recombinant plasmid pdg3 containing the entire mitochondrial (mt) genome of the fission yeast schizosaccharomyces pombe (s. pombe) cloned in the single bamhi-site of the e. coli plasmid pbr322 (del giudice 1981). by dna-rna hybridization it could be shown that the total mtdna sequence of the plasmid pdg3 was transcribed in the e. coli minicells. the cloned mtdna also directed the synthe ...	1983	6350830
expression and regulation of protein k, an escherichia coli k1 porin, in escherichia coli k-12.	using a modified lambda phage as a vector and a procedure developed in dr. c. schnaitman's laboratory, we have cloned the structural gene for protein k from an escherichia coli k1 strain to an e coli k-12 strain. the cloned inserts consist of two hindiii fragments, 4 kb and 6.5 kb in size. the protein produced by the insert is nearly identical to "authentic" protein k when chymotryptic peptides of 125i-labeled proteins are compared. protein k was found to respond to changes in the osmolarity of ...	1983	6373798
purification and characterization of reca protein from salmonella typhimurium.	reca protein was purified to homogeneity from salmonella typhimurium ta98 strain after induction of the cells by nalidixic acid. the purification was monitored with a radioimmune assay and involved a specific elution of the protein by atp from a single-stranded dna-cellulose column. from 240 liters of cell culture we obtained 40 mg of reca protein which was more than 98% pure. this protein exhibited the same molecular weight as measured on sodium dodecyl sulfate-polyacrylamide gel and the same i ...	1983	6219107
protein degradation in escherichia coli: the lon gene controls the stability of sula protein.	escherichia coli lon mutants are defective in the atp-dependent proteolysis of abnormal proteins. the mutants are also sensitive to ultraviolet light (uv) in that septation is inhibited after exposure to uv. sula mutations, isolated as suppressors of uv sensitivity unlinked to lon, do not affect proteolysis but allow septation to occur after dna damage. we have confirmed the hypothesis that the product of the sula gene is degraded by lon proteolysis. if sula (the product of sula) is a uv-inducib ...	1983	6300834
site-specific recombination by gin of bacteriophage mu: inversions and deletions.	a 3000-bp invertible segment in the dna of bacteriophage mu determines the host range of the phage. the inversion is catalyzed by the phage-coded protein gin; the recombination sites are short inverted repeats. gin protein is only made in low amounts by mu. to further investigate the gin-mediated recombination reaction a gin overproducing strain was constructed. the gin gene was cloned on a plasmid behind the pl-promotor of phage lambda. this results in a 100-fold higher inversion frequency of a ...	1983	6305017
use of lambda phasmids for deletion mapping of non-selectable markers cloned in plasmids.	a nonselectable gene carried on a poorly selectable recombinant plasmid has been physically mapped by deletion analysis. our method involved cloning the plasmid into a coliphage lambda vector and treating the recombinant phage with a chelator. virtually all particles surviving this treatment carried large deletions within the plasmid insert. further deletion analysis was done by inserting a selectable lambda sequence into one such deletion derivative and repeating the chelator selection. chelato ...	1983	6305773
formation of oligomeric structures from plasmid dna carrying cos lambda that is packaged into bacteriophage lambda heads.	plasmids that carry cos lambda, the region necessary for lambda phage packaging and that are as small as four kilobases in size can be packaged into lambda phage heads in head-to-tail tandem oligomeric structures. multimeric oligomers as large as undecamers have been detected. oligomer formation depends upon the products of red and gam of lambda, and the general recombination occurs between different plasmids that share homologous dna regions. the packaging efficiency of plasmids depends on its ...	1983	6217189
proteolytic processing of phage lambda tail protein gph: timing of the cleavage.	we describe a method for the rapid partial purification of intermediate structures of phage lambda tail assembly, using formaldehyde-fixed escherichia coli cells to precipitate tail-related structures. the purification depends on the specific interaction between the e. coli lambda receptor protein and lambda tail protein gpj. protein compositions of tail assembly intermediates were analyzed to determine when in the assembly sequence the minor tail protein gph is cleaved. gph joins the tail precu ...	1983	6220513
features of bacteriophage lambda: analysis of the complete nucleotide sequence.		1983	6222866
lambda mutation in the escherichia coli rho gene that inhibits the n protein activity of phage lambda.	certain escherichia coli rho mutations, exemplified by rho026, block the growth of phage lambda by interfering with phage gene expression. the phage gene n, whose product suppresses transcription termination, appears to be expressed normally in the mutants, and the functional stability of the n protein is not affected. our data suggest that these rho mutations allow transcription to terminate despite the presence of n. other e. coli mutants displaying a similar phenotype (nus(-)) fail to propaga ...	1983	6225121
gene transfer into animal cells after fusion with bacteriophage lambda-infected e coli protoplasts.		1983	6227013
escherichia coli plasmid vectors for high-level regulated expression of the bacteriophage lambda xis gene product.	the bacteriophage lambda xis protein is one of the proteins required for site-specific excisive recombination by which the lambda prophage is excised from the escherichia coli bacterial chromosome. we cloned the lambda xis gene under the control of several prokaryotic promoters to obtain a sufficient source of the protein for biochemical studies. our results demonstrate that e. coli lac promoter and lambda pl promoter fusions to the xis gene produce high levels of xis protein. induction of the e ...	1983	6229452
integration of viral dna into the genome of the adenovirus type 2-transformed hamster cell line he5 without loss or alteration of cellular nucleotides.	hamster cell line he5 has been established from primary lsh hamster embryo cells by transformation with adenovirus type 2 (ad2) (1). each cell contains two to three copies of integrated ad2 dna (2, 3). we cloned and sequenced the sites of junction between viral and cellular dnas. the terminal 10 and 8 nucleotides of ad2 dna were deleted at the left and right sites of junction, respectively. the integrated viral dna had an internal deletion between map units 35 and 82 on the ad2 genome. at the in ...	1983	6316259
plasmid vectors for the rapid isolation and transcriptional analysis of human beta-globin gene alleles.	we describe the construction and characterization of miniplasmid vectors that can be used to isolate and express normal and mutant alleles of the human beta-globin gene. these vectors, designated pi sv beta plasmids, contain a bacterial origin of replication and selectable marker, a 5'-flanking beta-globin dna fragment that can be used for recombination screening (seed, 1983), and simian virus 40 (sv40) sequences that allow accurate and efficient expression of the beta-globin gene transfected in ...	1983	6094959
expression of the replication region of phage lambda dna cloned into pbr322 in e. coli minicells.	replication region of bacteriophage lambda dna was cloned into pbr322 plasmid by the use of two restriction enzymes--psti and hindiii. the restriction analysis of four obtained plasmids revealed that lambda dna was cloned in both orientations. recombinant plasmids were transferred to the minicell-producing strain of e. coli and synthesis of the plasmid-mediated proteins was analysed by polyacrylamide-gel electrophoresis. all four recombinant plasmids produced lambda dna replication proteins po a ...	1982	6218724
solid-phase sequence analysis of polypeptides eluted from polyacrylamide gels. an aid to interpretation of dna sequences exemplified by the escherichia coli unc operon and bacteriophage lambda.	an approach to sequencing proteins by the solid-phase method combined with isolation of proteins and polypeptides by gel electrophoresis is described. mixtures of proteins or polypeptides resulting from digests are fractionated in the presence of dodecylsulphate in polyacrylamide gels. they are detected with coomassie blue, eluted, selectively reacted with porous glass derivatives and sequenced in their amino-terminal regions with the aid of a new microsequencer. alternatively they can be analys ...	1982	6210528
identity of a chi site of escherichia coli and chi recombinational hotspots of bacteriophage lambda.		1982	6210783
construction of recombinant lambda phages that carry the e. coli recb and recc genes.	a fragment of the e. coli chromosome including the recc gene has been cloned by in vitro recombinant dna techniques into a phage lambda vector to give the recombinant phage lambda drecc. this was used to derive the phage lambda drecbc by in vivo recombination. on lysogenisation of recb and recc mutants with lambda drecbc wild levels of uv-resistance and recbc dnase activity were restored. infection of e coli with lambda drecbc led to the synthesis of phage-coded proteins of 125 kilodaltons (kd) ...	1982	6211590
a comparison of the requirements for antitumour activity and antibacteriophage lambda activity for a series of non-intercalative dna-binding agents.	a series of non-intercalative dna-binding agents, comprising mainly bisquaternary ammonium heterocyclic compounds, has been found to inhibit strongly the production of bacteriophage lambda following its induction in escherichia coli. the inhibition is much greater than that found with a number of dna intercalating agents, including 9-aminoacridine, ethidium and daunorubicin. the inhibition correlated significantly with antitumour effect, as measured in a life extension assay with l1210 leukaemia ...	1982	6212240
purification of the bacteriophage lambda xis gene product required for lambda excisive recombination.	excision of the lambda prophage from the chromosome of its escherichia coli host requires the products of the two viral genes int and xis. this paper reports a purification of the lambda xis gene product using a complementation assay in which functional xis must be added to purified int and an e. coli-derived host factor extract. excisive recombination between a left (attl) and right (attr) prophage attachment site cloned on the same plasmid dna substrate occurred efficiently under these conditi ...	1982	6213611
cleavage of lambda repressor and synthesis of reca protein induced by transferred uv-damaged f sex factor.	transfer of a uv-damaged f sex factor to a recipient lambda lysogen induces prophage lambda development. under these conditions reca protein synthesis was induced and lambda repressor cleaved, as observed upon direct induction, that is, when the recipient lambda lysogen was directly exposed to uv-light. the efficiency of induction of reca protein synthesis in recipient bacteria which had received an irradiated f-lac factor was about 80% of that measured upon direct induction. we observed the sim ...	1982	6213837
analysis of lambda insertions in the fucose utilization region of escherichia coli k-12: use of lambda fuc and lambda arga transducing bacteriophages to partially order the fucose utilization genes.	escherichia coli k-12 strains have deletions for the normal lambda integration site were lysogenized with bacteriophage lambda at a site within the l-fucose utilization system (fuc). the frequency of lambda integration at this site is approximately 2 x 10(-8) to 5 x 10(-7). studies of the lytic properties of these strains indicated very infrequent cell lysis with a relatively low phage burst size. transductional ability of the phage lysates was found to be normal, comparable to that found in con ...	1982	6214544
mutation and w-reactivation of lambda phage by mitomycin c in the excision-defective escherichia coli.		1982	6216402
transcriptional termination sites in the b2 region of bacteriophage lambda that are unresponsive to antitermination.	a bacteriophage lambda cloning vector carrying the trp/lacw205 substitution is described. the vector facilitates the fusion in vitro of genetic control signals to the lacz structural gene of escherichia coli. this system was used to define transcriptional termination sites in the lambda b2 region. this region contains termination sites that are unresponsive to the lambda antiterminating proteins pq and pn.	1982	6285144
molecular cloning and amplification of the gene for thymidylate synthetase of e. coli.	the thya gene of escherichia coli, which directs the synthesis of the enzyme thymidylate synthetase, has been subcloned from a recombinant lambda phage (hickson et al., 1982) into the multicopy plasmid pbr325 to give the plasmid ppe245. to identify the thya gene product, the transposon tn1000 was inserted into ppe245 and derivative plasmids isolated that were no longer able to complement thya mutations. when proteins synthesised by these plasmids and by ppe245 were labelled and analysed on sds-p ...	1982	6290329
abelson murine leukemia virus: structural requirements for transforming gene function.	the integrated abelson murine leukemia virus (a-mulv) genome cloned in bacteriophage lambda gtwes.lambda b was used to localize viral genetic sequences required for transformation. comparison of the biological activity of cloned a-mulv genomic and subgenomic fragments showed that subgenomic clones that lacked the 5' long terminal repeat and adjoining sequences (300 base pairs downstream of the repeat) were not biologically active. in contrast, subgenomic clones that lacked the 3' long terminal r ...	1982	6291048
escherichia coli plasmid vectors containing synthetic translational initiation sequences and ribosome binding sites fused with the lacz gene.	the construction of a series of escherichia coli plasmid vectors suitable for assaying the effects of gene control signals fused with the e. coli lacz gene is reported. a synthetic deoxyoligonucleotide dodecamer 5'-catgaattcatg gtacttaagtac-5' containing two translation initiation codons (atg) separated by an ecori site was ligated with a lacz gene derivative which lacks the codons for the first eight amino acids in plasmid pmc1403 (casadaban et al., 1980). two ribosome-binding sequences were sy ...	1982	6293930
localization of the metjblf gene cluster of escherichia coli in lambda met transducing phage.	the position of the metjblf gene cluster in the transducing phage lambda met102 was determined by ligation of its leftmost ecori fragment (102-1) to the lambda bcdef (nin5) ecori fragment of lambda gtl (lambda bc) and characterization of the resultant recombinant phage. the new transducing phage carries about 6kb of bacterial dna which contains the entire met gene cluster including the promoter of its rightmost member metf. reasonable estimates of the coding capacity required for the four genes ...	1982	6294471
probing cii and hima action at the integrase promoter pi of bacteriophage lambda.	plasmids were constructed to supply cii-coded protein for activation of the phage promoter pi. using a fusion which expresses lacz from pi. we can accurately follow activation of pi without having to assay int activity in vivo. a large excess of cii protein compared to a normal lytic infection stimulates lacz expression about 10-fold over the basal level. the int-c226 constitutive allele of pi is not further activated by cii even though its level of lacz expression is less than the maximal cii-a ...	1982	6295882
site-specific dna condensation and pairing mediated by the int protein of bacteriophage lambda.	the int protein of bacteriophage lambda catalyzes the site-specific integrative recombination that inserts lambda dna into the host chromosome. the attachment site region of lambda dna required for this reaction spans 230 base pairs and includes four separable binding sites for int protein. we have used the electron microscope to determine the functional consequences of the interaction of int with its multiple binding sites. we find that int condenses a 230-base pair segment of dna into a compac ...	1982	6310548
cloning of colicin e1 tolerant tolc (mtcb) gene of escherichia coli k12 and identification of its gene product.	a mutation in the tolc(mtcb) gene of escherichia coli k12 results in increased sensitivity to sodium dodecylsulfate (sds), sodium deoxycholate, basic dyes, mitomycin c, and bleomycin, and makes the cell tolerant to the killing action of colicin e1. from lysogens with lambda ci857s7 integrated at a secondary attachment site, a transducing phage (lambda dtolc+) that transduces a tolc recipient to sds resistance was isolated. a recombinant dna molecule was constructed in vitro from plasmid pbr322 a ...	1982	6219270
identification of sequences necessary for packaging dna into lambda phage heads.	several species of dna molecules are packaged into lambda phage heads if they carry the region around the cohesive end site of lambda phage (cos lambda). the minimal functional sequence around cos lambda needed for packaging was examined by cloning in pbr322. the results showed that the minimal region contained 85 bp around cos lambda; 45 bp of the left arm of lambda phage and 40 bp of the right arm. a 75-bp region located to the right of the minimal region seems to enhance packaging. a 223-bp f ...	1982	6299893
studies on the association of e. coli phage lambda dna and the host chromosome: lack of a role of membranes.		1982	6461964
a system for genetic analysis in gene lamb: first results with lambda-resistant tight mutants.	we describe a system for genetic analysis in gene lamb. it consists of a phage which allows mapping, complementation and sequencing studies of lamb mutations and of the sequence of gene lamb. we present results obtained with this system for a set of mutations conferring tight resistant to phage lambda. this leads to a first identification of three residues in the lamb protein which are important for adsorption of phage lambda h+. residues 151 and 382 are important for reversible adsorption while ...	1982	6462090
expression of pyruvate formate-lyase of escherichia coli from the cloned structural gene.	it is shown here that a plasmid (p29) derived from the transducing phage lambda aspc2 (christiansen and pedersen 1981) codes for pyruvate formate-lyase. the identity of the 80 kilodaltons (kd) gene product of plasmid p29 with the pyruvate formate-lyase polypeptide was proven (i) by co-migration of the gene product expressed in the maxicell system with purified enzyme on o'farrell gels, and (ii) by comparison of the peptide maps obtained from limited proteolysis. in vivo the 80 kd form of the enz ...	1982	6758723
open reading frame cloning: identification, cloning, and expression of open reading frame dna.	a plasmid was constructed that facilitates the cloning and expression of open reading frame dna. a dna fragment containing a bacterial promoter and the amino terminus of the ci gene of bacteriophage lambda was fused to an amino-terminally deleted version of the lacz gene. an appropriate cloning site was inserted between these two fragments such that a frameshift mutation was introduced upstream of the lacz-encoding dna. this cloning vehicle produces a relatively low level of beta-galactosidase a ...	1982	6815653
extensive sequence homology in the dna coding for elongation factor tu from escherichia coli and the chlamydomonas reinhardtii chloroplast.	considerable dna sequence homology can be detected between the escherichia coli genes coding for translational components and chlamydomonas reinhardtii chloroplast dna. labeled chloroplast dna was found to hybridize to restriction fragments of the transducing phage lambda fus3 that code for elongation factor tu. the chloroplast probe also reacts with fragments coding for ribosomal proteins carried by this phage. the region homologous to the elongation factor genes was located on the physical map ...	1982	7048316
klebsiella and enterobacter strains derived from hospital infections. ii. occurrence and characterization of r-, lac- and col- plasmids and their clinical-epidemiological significance.	a total of 269 hospital klebsiella strains and 103 hospital enterobacter strains showed 34 and 10 different antibiotic resistance patterns, respectively. among multiple resistant klebsiella and enterobacter strains the ap sm cm tc resistance pattern was the most frequent (k. aerogenes). antibiotic resistant strains carried r-plasmids in 27.5%. the presence of r-plasmids was demonstrable in 2.9% of single antibiotic resistant, in 12.8% of double antibiotic resistant, and in 71.4% of multiple anti ...	1981	7257874
replication control and switch-off function as observed with a mini-f factor plasmid.	mini-f is a fragment of the f plasmid, consisting of 9,000 base pairs, which carries all of the genes and sites required for replicon maintenance and control. its copy number is one to two per chromosome. this plasmid is joined to cole1, whose copy number is 16 to 20. under normal circumstances the composite plasmid replication exhibited cole1 characteristics, maintaining a high copy number. however, when cole1 replication was inhibited by deoxyribonucleic acid polymerase i inactivation, its rep ...	1981	7021532
inducibility of a gene product required for uv and chemical mutagenesis in escherichia coli.	the product of the umuc gene is required for uv and chemical mutagenesis in escherichia coli. by the use of the mud(ap, lac) bacteriophage, we have obtained an operon fusion of the lac structural genes to the promoter/regulatory region of the umuc gene. the strain containing the umuc::mud(ap, lac) fusion was identified on the basis of its uv nonmutability. strains containing this putative null allele of umuc were (i) nonmutable by uv and other agents, (ii) slightly uv sensitive, and (iii) defici ...	1981	7029544
tif-1 mutation alters polynucleotide recognition by the reca protein of escherichia coli.	the requirements for polynucleotide-dependent hydrolysis of atp and for proteolytic cleavage of phage lambda repressor have been examined for both the wild-type (reca+ protein) and the tif-1 mutant form [tif(reca) protein] of the reca gene product. the reca+ and tif(reca) proteins catalyze both reactions in the presence of long single-stranded dnas or certain deoxyhomopolymers. however, short oligonucleotides [(dt)12, (da)14] stimulate neither the protease nor the atpase activities of the reca+ ...	1981	7031642
sos induction and autoregulation of the hima gene for site-specific recombination in escherichia coli.	the hima gene of escherichia coli controls the lysogenization of bacteriophage lambda at the level of catalysis of site-specific recombination and expression of the lambda int and ci genes required for lysogenic development. we have analyzed the regulation of hima by two methods: (i) beta-galactosidase synthesis from a lacz gene inserted into the hima gene and (ii) detection of radioactive hima protein after fractionation by two-dimensional gel electrophoresis. we find that hima- mutations produ ...	1981	6796964
rna splicing mutation in an aberrantly rearranged immunoglobulin lambda i gene.	the mouse cell line mopc 315 is an iga (lambda ii)-producing myeloma. we have studied a derivative of mopc 315 that secretes normal lambda ii chains but no heavy chain. this derivative, mopc 315-26, was found to contain a rearranged lambda i gene in addition to a rearranged lambda ii gene. the rearranged lambda i gene was cloned into bacteriophage lambda dna and its structure was studied. the lambda i gene was found to have arisen by an aberrant recombination event that resulted in a single base ...	1981	6171827
[low transfecting efficiency of phage lambda ring chromosomes and their fragments formed by membrane nucleases].	transfection efficiency of a number of lambda dna samples differing in ring to linear molecules ratio was determined. graphic extrapolation to the zero content of linear molecules showed that efficiency of ring molecules did not exceed 5% of that of linear molecules. probably, this difference is caused by more fast penetration of linear molecules into the cell and, therefore, by lower probability of their degradation by cell wall nucleases. fragments of both ring and linear molecules formed by c ...	1981	6453042
evidence that ribosomal protein s10 participates in control of transcription termination.	we report the isolation of an escherichia coli k-12 strain with a mutation, nuse71, that results in a change in ribosomal protein s10. phage lambda fails to grow in hosts carrying the nuse71 mutation because the lambda n gene product is not active. the n product regulates phage gene expression by altering transcription complexes so that they can overcome termination barriers. this suggests that a ribosomal protein is involved in antitermination of transcription.	1981	6453343
purification of bacteriophage lambda o protein that specifically binds to the origin of replication.	by means of a nitrocellulose filter binding assay, dna binding activities among proteins fractionated from extracts of escherichia coli carrying lambda dv have been surveyed. an activity was found that binds specifically to a fragment of 164 base pairs that specifies the lambda replication origin (lambda ori). this activity was not detected in an extract of cells not carrying the lambda dv plasmid. the activity was detected in extracts of cells carrying a hybrid plasmid in which the entire lambd ...	1981	6454055
arrangement of bacteriophage lambda receptor protein (lamb) in the cell surface of escherichia coli: a reconstitution study.	the lamb protein purified in a solution of sodium dodecyl sulfate was assembled into an ordered hexagonal lattice structure with a lattice constant of about 7.8 nm in the presence of lipopolysaccharide. the lamb alone formed aggregates with some lattice structure. however, the regularity of the lattice was only maintained within a very small area. an ordered hexagonal lattice was also formed when the wild-type lipopolysaccharide was replaced by heptoseless lipopolysaccharide, lipid a, and even f ...	1981	6455415
fusion of the lac genes to the promotor for the cytidine deaminase gene of escherichia coli k-12.	phage mu has been inserted into the structural gene for cytidine deaminase (cdd). by the use of phage lambda (lac, mu) the promoter for the cdd gene has been fused to lacz. in these strains lacz expression is regulated by the cytr repressor protein and is therefore induced by cytidine. the fusion strains were used for the isolation of cddo mutants. plaque forming lambda phages carrying the different cdd-lacz fusions were isolated. studies of the cdd-mu strains showed that the cdd gene is transcr ...	1981	6455590
structure and function of the phage lambda att site: size, int-binding sites, and location of the crossover point.		1981	6457725
effects of recb21, recf143, and uvrd152 on recombination in lambda bacteriophage-prophage and hfr by f- crosses.	the effects of the mutation pairs recb21 recf143 and recb21 uvrd152 on the frequency of genetic recombination were investigated in lambda phage-prophage crosses under homoimmune conditions. to prevent recombinants from being formed by the phage red system, these experiments were performed with phages and prophages carrying red and gam mutations. both spontaneous and damage-induced recombination was measured, the phages being either undamaged or treated with trimethylpsoralen and 360-nm light to ...	1981	6457825
preferential cleavage of phage lambda repressor monomers by reca protease.		1981	6457991
general method for fine mapping of the escherichia coli k-12 lamb gene: localization of missense mutations affecting bacteriophage lambda adsorption.	lamb is the structural gene for the bacteriophage lambda receptor, a multifunctional protein located in the outer membrane of escherichia coli k-12. we present a method for deletion mapping of any lamb mutations with a recognizable pheno-type. this method involves a transducing phage constructed by in vitro recombination which can also be used for complementation, deoxyribonucleic acid sequence, and in vitro protein synthesis studies with the mutated lamb gene. using this method, we mapped 18 la ...	1981	6458595
direction of bacteriophage lambda dna replication in a thymine requiring escherichia coli k-12 strain. effect of thymidine concentration.	the direction of replication was established for the first round of bacteriophage lambda dna replication in thymine requiring e. coli k-12 cells exposed to different concentrations of thymidine. it was found that a dramatic decrease in the proportion of bidirectionally replicating molecules followed a decrease in the concentration of thymidine. moreover, the rightward mode of replication appears to be exclusively favored in unidirectionally replicating molecules found at low concentrations of th ...	1981	6460985
plasmid vectors capable of transferring large dna fragments to yeast.	we have constructed several cloning vectors which can be used in vitro packaging and yeast transformation. these plasmids have been designed for the convenient cloning of large segments of dna and their transfer to yeast. they contain bacterial plasmid dna sequences for replication and selection in escherichia coli, yeast 2-microns plasmid dna sequences or chromosomal replicators and yeast markers necessary for replication and selection in yeast, and the cohesive ends of bacteriophage lambda whi ...	1981	6299664
the nus mutations affect transcription termination in escherichia coli.	the nusa1 and nusb5 mutations in a partial suppression of polarity and thus transcription termination in escherichia coli. as these mutations block the transcription antitermination activity of bacteriophage lambda n gene product, they paradoxically seem to enhance transcription termination at phage termination sites. the rho mutation hdf026 displays almost identical properties. the observations suggest that the nusa and nusb gene products may act as termination factors analogous to rho protein.	1981	6265784
overproduction of the ecorii endonuclease and methylase by escherichia coli strains carrying recombinant plasmids constructed in vitro.	recombinant dna molecules were constructed from the plasmid pil203 and the ecori-fragment of n3 plasmid containing ecorii endonuclease and methylase genes and also a gene for resistance to sulfanilamide. the pil203 plasmid, used as a vector, consisted of the bam hi-ecori-fragment of the plasmid pbr322 conferring resistance to ampicillin and the bam hi-ecori-fragment of lambda phage containing promoters, a thermosensitive mutation in the ci gene and a suppressible amber mutation in the cro gene. ...	1981	6266480
isolation of beta-globin-related genes from a human cosmid library.	a human gene library was constructed using an improved cloning technique for cosmid vectors. human placental dna was partially digested with restriction endonuclease mboi; size-fractionated and ligated to bamhi-cut and phosphatase-treated cosmid vector pjb8. after packaging in lambda phage particles, the recombinant dna was transduced into escherichia coli 1400 or hb101 followed by selection on ampicillin for recombinant e. coli. 150 000 recombinant-dna-containing colonies were screened for the ...	1981	6266915
cosmid cloning and transposon mutagenesis in salmonella typhimurium using phage lambda vehicles.	we have constructed a strain of salmonella typhimurium which contains the malb region from escherichia coli and carries the bacteriophage lambda receptor protein in its outer membrane. phage lambda adsorbs to this strain but cannot grow, thus providing a very useful system for transposon mutagenesis of s. typhimurium using lambda vehicles carrying transposons. this system can also be used for cosmid cloning.	1981	6268936
[artificial tn2551 transposon with replicative properties].	a hybrid plasmid, pbe10 was constructed. it consists of dnas of rsf2124 (cole1 :: tn3) plasmid and pub110 plasmid of staphylococcus aureus. the latter can be stably maintained in bacillus subtilis. bamhi cleaved pub110 was introduced into the bamhi site of transposon tn3 and the resulting enlarged tn3 (tn2551) was transposed from pbe10 onto phage lambda and than to pmb9 (tc) and rsf1010(sm su) plasmids. restriction and heteroduplex analysis of pmb9 :: tn2551(pbe21) and rsf1010 :: :: tn2551(pbe32 ...	1981	6273258
beta protein of bacteriophage lambda promotes renaturation of dna.	the protein encoded by the red beta gene of bacteriophage lambda was found to promote reannealing of complementary single strands of dna. reannealing activity was optimal at ph 6.0 and required a divalent cation. a threshold temperature of at least 15 degrees c was necessary in order to detect activity. the reaction was linear with time for about 20 min, but the extent of reaction was dependent upon the amount of beta protein added. reannealing of complementary single strands was confirmed by me ...	1981	6273399
construction and characterization of a tufa-lacz fusion coding for an e. coli ef-tu-beta-galactosidase chimeric protein.	a new phage lambda cloning vector was constructed that has a single ecori site upstream from weakly expressed laci-z gene isolated by müller-hill and kania (1974). an ecori fragment containing the complete tufa gene of e. coli was cloned on the vector and the recombinant phage was crossed into the str operon that has tufa as its last gene. subsequent selection gave rise to a tufa-lacz fusion that codes for a chimeric peptide. the fused peptide has a molecular weight of 148,000 and contains 40% o ...	1981	6276696
[bacteriophage lambda integration into host chromosome (biochemistry of int protein and pleiotropic effects of host factors) (author's transl)].		1980	6243784
construction and properties of a cole1::tn3-cos lambda plasmid for determining rna polymerase binding sites on cole1 and tn3.	to determine the location of the rna polymerase binding sites on the cole1 plasmid and tn3 transposon, a special hybrid cole1::tn3-cos lambda molecule was constructed which contains the left arm of phage lambda dna and the right lambda terminal fragment. this permits orienting cole1 molecules, since the rna polymerase binding pattern of these two lambda fragments are known to be distinct. cole1 dna contains seven binding sites and tn3 binds three rna polymerases, with some of the latter probably ...	1980	6247244
cloning of the replication gene o of e. coli bacteriophage lambda and its expression under the control of the lac promoter.	the expression of the replication gene o of bacteriophage lambda was put under the control of the lac promoter-operator region integrated into the pbr322 cloning vehicle. the new plasmid pkk104 was introduced into minicells and the o gene induced by isopropyl-beta-thiogalactoside (iptg). the o protein could be identified as a major component in extracts from these cells, in association with the cell membrane fractions. the molecular weight of the o protein in sds gels is about 33 000, and it is ...	1980	6254838
bacteriophage lambda cloning vehicles for studies of genetic recombination.	a pair of bacteriophage lambda cloning vehicles has been constructed for use in studies of genetic recombination. these phages, lambda rva and lambda rvb, have the following properties: (1) each vector has a single hindiii site in the immunity region, at which segments of dna can be inserted. (2) these hindiii sites are flanked by selectable markers with the following phenotypes: spi+/- (fec+/-) to the left, and imm lambda or imm434 to the right. (3) there is essentially no sequence homology bet ...	1980	6254844
the ral gene of phage lambda. i. identification of a non-essential gene that modulates restriction and modification in e. coli.	host controlled restriction in escherichia coli can be relieved by pre-infecting restricting cells with modified lambda helper phages. this process, in which intact unmodified phage genomes are allowed to escape restriction attack, is mediated by a newly identified lambda function called ral. the ral gene has been located by deletion mapping between ciii and n. efficient expression of the ral gene requires the product of the regulator gene n. polyacrylamide gel analysis of the lambda proteins sp ...	1980	6256607
cloning of the exonuclease iii gene of escherichia coli.	overproducers of exonuclease iii (exo iii) were found within a colony bank containing cole1-escherichia coli hybrid plasmids. through the enzymatic ligation of restriction enzyme fragments, the exo iii gene, xth, was transferred to a thermoinducible, integration-proficient lambda phage and to a chimeric cole1-lambda plasmid that was thermoinducible for lambda-directed dna replication. transfer of the xth gene was facilitated by a technique involving prior selection for tn5 insertions into plasmi ...	1980	6260569
l factor that is required for beta-galactosidase synthesis is the nusa gene product involved in transcription termination.	the dna-dependent in vitro synthesis of escherichia coli beta-galactosidase requires the presence of a soluble protein referred to as l factor [kung, h., spears, c. & weissbach, h. (1975) j. biol. chem. 250, 1556-1562]. in the present study, comparison of physical, immunological, and biological properties shows that l factor is the product of the e. coli nusa gene. the nusa gene product is known to interact with bacteriophage lambda n gene protein and to prevent premature termination of transcr ...	1980	6154941
studies on the e. coli gronb (nusb) gene which affects bacteriophage lambda n gene function.	escherichia coli mutants, called gronb, which block the growth of bacteriophage lambda at the level of action of the gene n product, have been isolated as survivors at 42 degrees c of bacteria carrying a) the defective prophage lambda bio11 i lambda ci857 delta h1 or b) the pcr1 plasmid containing the ecori immunity fragment of phage lambda ci857. in addition, gronb bacterial mutants have been isolated at 37 degrees c, as large colony formers in the presence of lambda i lambda ci h434, lambda i ...	1980	6161293
expression of prokaryotic genes inserted into cole1 and pvh51 plasmids.	one of the dna fragments obtained from ecori digests of guaa-transducing lambda phage dna contains the intact bacterial guaa gene at its one end and the lambda phage r gene at the other end. this dna fragment, named coslambda-guaa, does not contain promoter-operator regions of the gua operon and of the lambda phage r gene, coslambda-guaa dna fragments were inserted in two different orientations into respective dnas of ecori-cleaved cole1 and pvh151 (= mini cole1). mitomycin c stimulated the guaa ...	1980	6444526
purification and characterization of the integration protein specified by bacteriophage lambda.		1980	6444632
[adsorption of lambda phage dna onto escherichia coli cells treated with ca2+ ions and onto frozen--thawed bacteria].	the study of the biologically active tritium-labeled phage lambda dna adosrption on ca2+-treated and frozen--thawed e. coli cells showed the absence of a correlation between the adsorption level and transfection efficiency. thus the infectious phage lambda dna adsorption level does not change, while frozing--thawing of e. coli cells but it increases ten-fold when treating the cells with ca2+ in ice, the transfection efficiency level with this dna being equal for both types of recipients.	1980	6446330
induction of prophage lambda without amplification of reca protein.	the requirement for amplified synthesis of reca protein in the uv-promoted induction of coliphage lambda was studied. we confirmed that a low concentration of rifampicin inhibited specifically the increased synthesis of reca protein after an inducing treatment (satta and pardee, 1978). under these conditions, using an optimal dose of uv, e. coli lysogens were induced, producing active phage. the drug delayed the onset of induction and with increasing concentrations affected the yield of phage, b ...	1980	6446647
pseudovirulent mutants of lambda b221poricasna resulting from mutations in or near oric, the e. coli origin of dna replication.	mutants of the specialized transducing phage lambda b221poricasna have been isolated which form plaques on lambda lysogens. genetic and physical evidence is presented to show that the mutations responsible for the pseudovirulent phenotype map in or near oric, the origin of chromosomal replication in escherichia coli.	1980	6446650
the phage promoter responsible for the expression of the inserted beta-galactosidase gene in bacteriophage lambda plac5.	the lac transducing phage, lambda plac5, carries a segment of the e. coli lac operon on the left side of the b2 region of the lambda phage. in the absence of additional cyclic amp, beta-galactosidase can only be expressed from the phage promoter, and the expression of the inserted lac promoter is suppressed. this phage promoter responsible for beta-galactosidase synthesis is shown to be under the control of the ci and n gene products; however, the repressive action of the cro gene product at hig ...	1980	6446653
transcription antitermination by bacteriophage lambda n gene product.		1980	6447798
promoter for the establishment of repressor synthesis in bacteriophage lambda.	transcription of the lambda repressor gene (ci) is positively regulated by the phage-encoded proteins cii and ciii. we have isolated and characterized the 5'-terminal region of this rna and shown that it originates at a promoter (pe) located between genes cro and cii. the dna sequence of this promoter shows little homology to other known promoters. initiation of transcription from pe is abolished by the cis-dominant mutations cy; these mutations alter the "-10" and "-35" regions of the promoter. ...	1980	6447872
[genetic properties of phage lambda recombinant molecules. the effect of an inserted fragment on the hybrid yield and recombination with respect to the h- and vir-markers].	the yield of mature phages in lambda gt-lambda dm34, lambda gt-lambda dm225 and lambda gt-lambda dm234 is 2, 8 and 8 times lower (respectively) as compared to the wild type. the output of hvir-recombinants in lambda gt-lambda dm225 is more than an order lower as compared to the other phages. as distinct from two other hybrids, the decrease in lambda gt-lamba dm225 yield cannot be explained by red-genotype and a shortening of the phage genome as well as the decrease in the output hvir-recombinant ...	1980	6449098
in vitro comparison of initiation properties of bacteriophage lambda wild-type pr and x3 mutant promoters.	the in vitro initiation properties of the pr promoter of bacteriophage lambda and of a pr mutant, x3, were compared. using the abortive initiation reaction, we measured the lags in the approach to a final steady-state rate when dinucleotide synthesis was initiated with rna polymerase. these lags corresponded to the average times required for the formation of transcriptionally active open complexes. by measuring the lags at different rna polymerase concentrations, we could separate open complex f ...	1980	6450417