Publications

cloning and expression of a tylosin resistance gene from a tylosin-producing strain of streptomyces fradiae.	a gene conferring high-level resistance to tylosin in streptomyces lividans and streptomyces griseofuscus was cloned from a tylosin-producing strain of streptomyces fradiae. the tylosin-resistance (tylr) gene (tlra) was isolated on five overlapping dna fragments which contained a common 2.6 kb kpni fragment. the kpni fragment contained all of the information required for the expression of the tylr phenotype in s. lividans and s. griseofuscus. southern hybridization indicated that the sequence co ...	1986	3020383
structure and function of the repressor of bacteriophage lambda. iii. molecular cloning of the high-affinity mutant ci gene of lambda and studies of the properties of the clones.	the high-affinity mutant ci gene of lambda ciha (nag et al. 1984) was cloned in the multicopy plasmid pbr322. in the resulting plasmid, pmd 102, a lacuv5 promoter was inserted giving the lacuv5-ciha fusion plasmid pmd 205. bacteria carrying pmd 102 and pmd 205 contain 2.5 and 15 times, respectively, the level of repressor in a monolysogen of lambda ciha. results of the study of certain properties of the bacteria carrying these plasmids suggest that the ha repressor also has a higher affinity for ...	1986	3020384
specialized nucleoprotein structures at the origin of replication of bacteriophage lambda: localized unwinding of duplex dna by a six-protein reaction.	the o protein of bacteriophage lambda localizes the initiation of dna replication to a unique site on the lambda genome, ori lambda. by means of electron microscopy, we infer that the binding of o to ori lambda initiates a series of protein addition and transfer reactions that culminate in localized unwinding of the origin dna, generating a prepriming structure for the initiation of dna replication. we can define three stages of this prepriming reaction, the first two of which we have characteri ...	1986	3020552
[cleavage of dna adsorbed on the surface of phospholipid membranes by restrictases type ii].	cleavage of phage lambda dna by restriction endonucleases in the presence of model phosphatidylcholine membranes was studied. bsp1, pst1 and bam h1 were found to cleave dna under these conditions to a considerably decreased extent. this effect does not result from irreversible inactivation of the enzymes or their direct interaction with the membranes. the most probable explanation of the membrane inhibitory effect is the change of dna substrate properties resulting from its mg2+-mediated binding ...	1986	3021244
charomids: cosmid vectors for efficient cloning and mapping of large or small restriction fragments.	charomids are cosmid vectors up to 52 kilobases (kb) long, bearing 1-23 copies of a 2-kb spacer fragment linked in head-to-tail tandem arrays. like cosmids and lambda phage, charomids can be packaged in vitro for efficient introduction into bacteria. charomids contain a polylinker with nine unique restriction sites for cloning and can be used without preparing vector arms. using a charomid of appropriate size, one can clone inserts of any size up to 45 kb. for example, charomid 9-36 (9 cloning s ...	1986	3022301
[insertion of the cos-site into dna of phage m13 and its packing in proteins of phage lambda].	the cos-site of lambda phage from phc79 cosmide is transferred to dna from m13 mp18 phage. the recombinant dna thus obtained (mc18) is efficiently packaged into lambda proteins in vitro. the bamhi-hindiii fragment of pgp588 (a pbr322 derivatives containing fragment of human dna) is subcloned into mc18. although this pgp588 fragment contains numerous alu repeats, no essential rearrangements of the insert were revealed. the efficiency infection by recombinant dna packaged with lambda proteins is a ...	1986	3022756
extensive sequence homology of the goldfish ras gene to mammalian ras genes.	we cloned ras-related sequences from goldfish genomic libraries constructed as recombinants using the lambda phage. restriction enzyme mapping of the clones obtained revealed three kinds of ras-related sequences among approximately 350,000 genomic clones. one of these clones was partially sequenced. comparison with the nucleotide sequences of mammalian ras genes showed that the determined sequences covered the predicted amino acid coding regions and parts of the intervening regions. the predicte ...	1986	3023161
regulatory defects of a conditionally lethal nusats mutant of escherichia coli. positive and negative modulator roles of nusa protein in vivo.	previous studies have attributed two activities to the nusa protein of escherichia coli; namely, termination and antitermination of transcription. to examine these activities, we isolated a temperature-sensitive mutant of the nusa gene (nusats11). the mutant cells produce a thermolabile nusa protein and grow at 32 degrees c, but not at 42 degrees c. at 42 degrees c, nusats11 is recessive to nusa+ and nusa1, indicating the absence of its active gene product at that temperature. in the mutant, the ...	1986	3023618
transcription termination and processing sites in the bacteriophage lambda pl operon.	s1 nuclease mapping was performed on transcripts from the major leftward operon of the bacteriophage lambda in order to locate the 3' ends of stable rna species produced in vivo. the analysis was carried out on rna purified from either an induced lambda prophage or bacteria carrying a plasmid containing a large segment of lambda including the intact pl operon through the bet gene. the s1 nuclease mapping was performed on transcripts produced in the presence and the absence of the n antiterminati ...	1986	3023619
modification enhancement by the restriction alleviation protein (ral) of bacteriophage lambda.	the product of the lambda ral gene alleviates restriction and enhances modification by the escherichia coli k-12 restriction and modification system. an open reading frame (orf) located between genes n and ea10 has been assigned to the ral gene. we have cloned this orf in a plasmid where its transcription is controlled by a thermolabile lambda repressor. inactivation of the lambda repressor caused a 1000-fold reduction in k-specific restriction of unmodified lambda phage and a 100-fold increase ...	1986	3023633
introduction to recombinant dna.	this paper describes the current state of knowledge of methods for analysing gene structure and localization. illustrations are given of the preparation of complementary dna libraries and their screening by positive-negative selection, the use of synthetic oligodeoxynucleotides and the use of antibodies. analysis of the egf precursor is used as an example to show its close relationship to plasma membrane receptor and its homology with the ldl receptor. analysis of cloned genome dna by use of bac ...	1986	3023749
[characteristics of the formation and properties of pls plasmids resulting from deletions of bacteriophage lambda att80 (tn9) genome].		1986	3023978
the purification of the escherichia coli uvrabc incision system.	the uvra, uvrb and uvrc proteins of escherichia coli have been purified in good yields to homogeneity with rapid three- or four-step purification procedures. the cloned uvra and uvrb genes were placed under control of the e. coli bacteriophage lambda pl promoter for amplification of expression. expression of the uvrc gene could not be amplified by this strategy, however, subcloning of this gene into the replication-defective plasmid prlm24 led to significant overproduction of the uvrc protein. t ...	1986	3024108
cloning and sequence analysis of rat bone sialoprotein (osteopontin) cdna reveals an arg-gly-asp cell-binding sequence.	the primary structure of a bone-specific sialoprotein was deduced from cloned cdna. one of the cdna clones isolated from a rat osteosarcoma (ros 17/2.8) phage lambda gt11 library had a 1473-base-pair-long insert that encoded a protein with 317 amino acid residues. this cdna clone appears to represent the complete coding region of sialoprotein mrna, including a putative aug initiation codon and a signal peptide sequence. the amino acid sequence deduced from the cdna contains several ser-xaa-glu s ...	1986	3024151
human cholesterol side-chain cleavage enzyme, p450scc: cdna cloning, assignment of the gene to chromosome 15, and expression in the placenta.	conversion of cholesterol to pregnenolone is mediated by p450scc [cholesterol, reduced-adrenal-ferrodoxin: oxygen oxidoreductase (side-chain-cleaving), ec 1.14.15.67]. rna from several human adrenal samples was translated in vitro and immunoprecipitated with anti-bovine p450scc, indicating that p450scc mrna represents about 0.5% of human adrenal mrna in normal, hypertrophied, and malignant adrenals. a 1626-base-pair human adrenal p450scc cdna was cloned in bacteriophage lambda gt10. primer exten ...	1986	3024157
characterization of the topoisomerase ii-induced cleavage sites in the c-myc proto-oncogene. in vitro stimulation by the antitumoral intercalating drug mamsa.	in an attempt to get an insight into the activity of mamsa (a dna topoisomerase ii-mediated drug) on the human proto-oncogene c-myc, an in vitro system consisting of purified calf thymus dna topoisomerase ii and a c-myc dna inserted in lambda phage was utilized. the occurrence of discrete bands, detected by hybridization of southern blots with appropriate c-myc probes, indicated the presence of cleavage sites in the sole presence of dna topoisomerase ii. the band intensity increased in the prese ...	1986	3024649
cooperative dna binding by lambda integration protein--a key component of specificity.	quantitative analysis of nitrocellulose filter binding data by the method of clore, gronenborn and davies [(1982) j. mol. biol. 155, 447-466] has been used to show that lambda integration protein (int) exhibits cooperativity in binding to specific recognition sites within the attachment site region (lambda attp) of bacteriophage lambda dna. optimal values of the equilibrium constant obtained were 3.0(+/- 1.0) x 10(10) m-1 for the p' site using a model of three sites with equal affinity and 1.9(+ ...	1986	3024982
isolation and characterization of the gene for myosin light chain two of drosophila melanogaster.	a recombinant lambda-phage dna clone containing drosophila melanogaster sequences encoding the gene for myosin light chain (mlc) two has been isolated from a library of randomly sheared dna. the drosophila mlc2 gene is located in region 99e1-3 on the right arm of chromosome 3, several bands removed from the site reported for the other myosin light chain gene at 98b. the mlc2 sequence at 99e1-3 appears to encode all of the isoforms of drosophila mlc2. the polypeptide encoded at 99e was identified ...	1987	3025224
[different effect of mutations in escherichia coli k12 dna-genes on the transposition of tn5- and tn10-elements].	the effect of mutations in dnaa(dnaa46), dnag(dnag3), dnac (dnac1 and dnac2) and dnab genes on transposition of two transposons, tn5 and tn10, from bacteriophage lambda genome into the chromosome of host cells has been studied. transposition was performed at permissive temperatures for the mutant recipients. the mutations in dnaa, dnac, dnag genes were shown to decrease the transposition of tn10 for some orders of magnitude as compared with transposition registered in wild type cells. independen ...	1985	3025676
[deletion of the phage lambda att80 tn9 genome; the type of intramolecular recombination].	the intramolecular deletion-generating recombination which transforms lambda bacteriophage genomes into the plasmids (named pls) proved to be site-specific to a certain extent. using electron microscopy heteroduplex analysis three preferential sites for this recombination were found in seven independent pls isolates studied. att-sites were not registered to be involved in the formation of deletions in isolates studied. it was shown that recombination operating in our system was independent of th ...	1986	3025679
[effectiveness of incorporation of spermine-condensed viral dna into liposomes. interaction of liposomes with cells].	dna from bacteriophage lambda or monkey adenovirus type 7 have been condensed with spermine and included into three types of liposomes: ethereal, obtained by inverted phases technique and by ca++ fusion. compact dna form presents a tor with 100 nm external diameter and 430 nm width. it can be included into 100 nm or larger liposomes. total preparation contains 15-18% of the required liposomes. liposomal fractions with included dna were separated from empty liposomes by step gradient of ficoll 40 ...	1985	3025690
[cloning of pectate-lyase genes of erwinia chrysanthemi in escherichia coli cells].	erwinia chrysanthemi dna fragment digested by restriction endonuclease ecori and carrying the gene ec16 determining the synthesis of pectatelyase with rf 0.20 and mol. mass 40kd has been cloned in plasmid puc 9 plasmid in escherichia coli hb101 cells. three genes for pectatelyases of erwinia chrysanthemi ena49 have been cloned in vector phage lambda 47.1 in escherichia coli cells. two genes determining the synthesis of pectatelyases with rf 0.06 and 0.19 and mol. masses 40 kd and 39 kd have been ...	1986	3025699
[synthesis of aminoglycoside phosphotransferase is under control the pl-promoter of phage lambda].	the recombinant plasmid pkp145 pl has been constructed containing the gene for aminoglycosidephosphotransferase (apt). expression of the apt gene is under the control of lambda bacteriophage pl promoter. escherichia coli cells harbouring this plasmid synthesize apt in quantity up to 13-15% of the total cellular protein. the technique for isolation of apt from superproducing cells has been elaborated. preparations of the enzyme devoid of contaminating bacterial proteins have been obtained.	1986	3025708
structure and expression of the gene encoding the vasoactive intestinal peptide precursor.	the gene encoding the human vasoactive intestinal peptide (vip) and the histidine-methionine amide (phm-27) peptide hormone was isolated from lambda phage libraries. the human gene was found to be composed of seven exons spanning approximately 9 kilobase pairs. the first exon codes for an untranslated leader sequence, and the second exon codes for a putative signal peptide. dna sequences coding for the vip and phm-27 hormones are located in two different exons. southern blot analysis with genomi ...	1987	3025882
[effect of thiol-specific reagents on the activity of paei and paeii restrictases from pseudomonas aeruginosa].	5,5'-dithiobis-2-nitrobenzoic acid, n-ethylmaleimide, and parachloromercuribenzoate have been demonstrated to inhibit the activity of restrictases paei and paeii from ps. aeruginosa bacterial cells. restrictase paeii was more sensitive to the action of thiol-specific reagents, as compared to paei. the minimal concentration of reagents for sh-groups that completely inhibited the activity of restrictases paei and paeii was determined. the protective effect against the inhibitory action of 5,5'-dit ...	1986	3026511
nuclear reconstitution in vitro: stages of assembly around protein-free dna.	we have developed a cell-free system derived from xenopus eggs that reconstitutes nuclear structure around an added protein-free substrate (bacteriophage lambda dna). assembled nuclei are morphologically indistinguishable from normal eukaryotic nuclei: they are surrounded by a double membrane containing nuclear pores and are lined with a peripheral nuclear lamina. nuclear assembly involves discrete intermediate steps, including nucleosome assembly, scaffold assembly, and nuclear membrane and lam ...	1987	3026635
[plasmid vector for gene expression containing the temperature-dependent promoter of phage lambda p'r dna].		1986	3026761
in vitro packaging into phage t4 particles and specific recircularization of phage lambda dnas.	concatemeric phage lambda imm434 dna packaged in vitro into phage t4 particles produced plaques on a selective host. moreover, lambda dna containing a pbr322 derivative flanked by the lambda attl and attr sites could be specifically recircularized by excisive lambda recombination to yield the pbr322 derivative. a host deficient in generalized recombination and containing a defective lambda c its prophage which provided int and xis proteins was the recipient for this plasmid derivative carried by ...	1986	3026930
characterization of a cdna for human protein c inhibitor. a new member of the plasma serine protease inhibitor superfamily.	a cdna library in lambda-phage lambda gt11 containing dna inserts prepared from human liver mrna was screened with monoclonal antibodies to human protein c inhibitor. six positive clones were isolated from 6 x 10(6) phages and plaque purified. the cdna in the phage containing the largest insert, which hybridized to a dna probe prepared on the basis of the amino-terminal amino acid sequence of the mature inhibitor, was sequenced. this cdna insert contained 2106 base pairs coding for a 5'-noncodin ...	1987	3027058
regulation of expression of the gene for vitamin b12 receptor cloned on a multicopy plasmid in escherichia coli.	the btub gene of escherichia coli codes for a protein (btub) located in the outer membrane. btub is the receptor for vitamin b12 (cyanocobalamin). we have cloned the btub gene into puc8 using transposon tn5 as the marker to first isolate several parts of the relevant dna fragment from the specialized transducing phage lambda darg13. after reconstitution of the gene, tn5 was removed by selecting for spontaneous excision. the partial nucleotide sequence and transcriptional start of the btub gene w ...	1986	3027510
[the effect of the phage lambda ral gene on the level of synthesis of the ecok restriction endonuclease beta-subunit].	e. coli hsd genes were subcloned from lambda 642 (ral+) into lambda l47.1 vector (ral-after replacement). the influence of bacteriophage lambda ral gene on the expression efficiency of hsds kappa, hsdm kappa genes was investigated. it was shown, that its presence in vitro enhanced the synthesis of beta-subunit, hsdm gene product, and the increase of modification in vivo was observed. it is proposed that the increase of modification rate of lambda phage fully unmodified dna is connected with the ...	1986	3027537
[structural-functional organization of r-plasmid pbs222 with a broad range of bacterial hosts].	the broad host-range plasmid pbs222 is compatible with broad host-range plasmids of all known incompatibility groups and codes for tetracycline resistance. pbs222 is efficiently mobilized by inc p-1 plasmid rp4 and is also capable of conjugal transfer with low efficiency to different gramnegative microorganisms. the size of the plasmid (17.2 kb) has been determined and its physical map has been constructed. the plasmid harbours the unique sites for restriction endonucleases bglii, hindiii, hpai, ...	1986	3027554
a bacterial protein requirement for the bacteriophage lambda terminase reaction.	the bacteriophage lambda terminase enzyme cleaves the cohesive-end sites of lambda dna to yield the protruding 5'-termini of the mature molecule. in vitro, this endonucleolytic event requires a protein factor which has been isolated and purified from extracts of uninfected e. coli. the terminase host factor (thf) is a heat stable basic protein of m.w. approximately 22,000. the integration host factor (ihf) protein of e. coli can efficiently substitute for thf in the terminase reaction; however, ...	1986	3027664
characterization of the integration site of the cmv mtr in a tumor cell line.	previous studies have shown that a 558-bp fragment of human cytomegalovirus (cmv) dna contained within pcm4127 and designated cmv mtr can morphologically transform rodents cells in vitro. by cotransformation with pcm4127 and a plasmid conferring g418 resistance, psv2neo, morphologically transformed nih3t3 cell lines were isolated. dot blot hybridization indicated that approximately 30% of the transformants retained cmv sequences. two cell lines which retained viral dna were chosen for further st ...	1987	3027970
complete sequence and structure of the gene for human adenosine deaminase.	the nucleotide sequence of the human adenosine deaminase gene was determined. the gene was isolated in a series of overlapping lambda phage clones containing human germ line dna. a total of 36,741 base pairs were sequenced, including 32,040 base pairs from the transcription initiation site to the polyadenylation site, 3935 base pairs of 5'-flanking dna, and 766 base pairs of 3'-flanking dna. the gene contains 12 exons separated by 11 introns. the exons range in size from 62 to 325 base pairs whi ...	1986	3028473
the structure of the rat glutathione s-transferase p gene and related pseudogenes.	we have isolated the rat placental-type glutathione s-transferase (gst-p) gene from a lambda phage library using gst-p cdna clone, pgp5 (sugioka, y., kano, t., okuda, a., sakai, m., kitagawa, t., and muramatsu, m. (1985) nucleic acids res. 13, 6049-6057), as a probe. the rat gst-p gene is about 3 kilobase pairs long and contains 7 exons and 6 introns, encoding the same gst-p protein specified by pgp5. the cap site maps 70 nucleotides upstream from the translation initiation site. the canonical p ...	1987	3029128
human papillomavirus type 16 dna from a vulvar carcinoma in situ is present as head-to-tail dimeric episomes with a deletion in the non-coding region.	a number of genital cancer biopsy samples were screened for the presence of human papillomavirus type 16 (hpv-16) dna sequences. one of these samples (a vulvar carcinoma in situ) was found to contain more than 100 copies of hpv-16 dna sequences per cell. using this tumour dna, a genomic library was constructed in bacteriophage lambda and the library was screened for recombinant phage containing hpv-16 sequences. five recombinant phage clones were isolated and their dna was analysed by restrictio ...	1987	3029284
synthesis in escherichia coli and immunological characterization of a polypeptide containing the cleavage sites associated with trypsin enhancement of rotavirus sa11 infectivity.	about 45% of the rotavirus sa11 vp3 gene was inserted into a thermoinducible expression plasmid under the control of phage lambda pl promoter. the primary translation product predicted on the basis of the plasmid construction was a hybrid protein in which the 98 amino-terminal amino acids of phage ms2 polymerase were followed by amino acids 42 to 387 of the vp3 protein, which included the region containing the cleavage sites associated with trypsin enhancement of infectivity. on induction, a pol ...	1987	3029294
cloning of a human s-phase cell cycle gene: use of transient expression for screening.	we report here the cloning of a human cell cycle gene capable of complementing a temperature-sensitive (ts) s-phase cell cycle mutation in a chinese hamster cell line. cloning was performed as follows. a human genomic library in phage lambda containing 600,000 phages was screened with labeled cdna synthesized from an mrna fraction enriched for the specific cell cycle gene message. plaques containing dna inserts which hybridized to the cdna were picked, and their dnas were assayed for transient c ...	1987	3029567
[irreversible changes in phage dna after its protonation in solution and inside the virion detected by the transfection method].	the dependence of irreversible structural changes in phage lambda dna on the degree of its protonation in a solution and inside the virion has been found by measuring the transfection activity of bacteriophage. the different effect of ionic strength on ph-dependence of the irreversible changes in the structure of dna upon its protonation in a solution or in situ has been registered and explained. the insignificant shift of ph from neutral region value in 0.1 m nacl has resulted in a damaging eff ...	1986	3029576
the control of lambda dna terminase synthesis.	nu1 and a, the genes coding for bacteriophage lambda dna terminase, rank among the most poorly translated genes expressed in e. coli. to understand the reason for this low level of translation the genes were cloned into plasmids and their expression measured. in addition, the wild type dna sequences immediately preceding the genes were reduced and modified. it was found that the elements that control translation are contained in the 100 base pairs upstream from the initiation codon. interchangin ...	1987	3029667
characterization of the 5'-flanking region for the human fibrinogen beta gene.	to identify the possible regulatory sequences in the genetic expression of fibrinogen, a human genomic dna library raised in lambda embl 4 phage was screened using cdna probes coding for the a alpha, b beta and gamma chains of human fibrinogen. the entire fibrinogen locus was characterized and its organization analysed by means of hybridization and restriction mapping. among the clones identified, a single recombinant lambda phage contained the beta gene and its 5'- and 3'-flanking regions. a 1. ...	1987	3029722
the human growth hormone gene locus: structure, evolution, and allelic variations.	genomic clones containing the closely related genes for human growth hormone (hgh) and chorionic somatomammotropin (hcs) were obtained from genomic bacteriophage lambda and cosmid libraries. the entire gh/cs chromosomal locus was reconstructed utilizing overlapping restriction fragments characterized from the isolated clones. the hgh/hcs locus contains two gh genes and three cs genes spanning 48 kb of dna in the order: 5'-(hgh-1/hcs-5/hcs-1/hgh-2/hcs-2)-3', confirming analysis of cosmid clones o ...	1987	3030680
overproduction and large-scale preparation of deoxyuridine triphosphate nucleotidohydrolase from escherichia coli.	a recombinant plasmid, phw1, directing the overproduction of the enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dutpase, ec 3.6.1.23) from escherichia coli has been constructed. a 1900-base dna fragment carrying the structural gene for the enzyme (dut) has been recloned into a runaway replication vector that also carries the strong leftward promoter (pl) of bacteriophage lambda. upon temperature shift, an e. coli strain carrying the new plasmid gives an increase in dutpase activity of ...	1987	3030754
[molecular causes of thalassemia. iv. cloning of the beta-globin gene in a patient with beta-thalassemia from azerbaijan and determination of the point mutation in a minor intron].	on the basis of dna from a beta-thalassemic patient, human gene library has been obtained using bacteriophage lambda embl4 as a vector. the recombinants contain human dna insertions of 15 to 20 kb. the lambda a1 beta 1 clone has been isolated by the method of hybridization of phage plaque replicas to the hhai fragment of jw102 plasmid containing human beta-globin cdna. restriction mapping revealed the presence of a 22 kb human dna fragment comprising a portion of the beta-globin gene system. sub ...	1987	3030889
directional control of site-specific recombination by bacteriophage lambda. evidence that a binding site for int protein far from the crossover point is required for integrative but not excisive recombination.	phage lambda controls its integration and excision by differential catalysis of the forward and reverse reactions. the lambda int protein is required for both directions, but xis for excision only. to investigate the substrate requirements for directional control, we have characterized two mutations of the phage attachment site that are defective in integrative but not excisive recombination. both of these mutations produce the same base change in the p'3 binding site for int protein 79 base-pai ...	1986	3031315
[various characteristics of the anti-restriction mechanism in bacteriophage t5].	bacteriophage t5 is not confined by the restriction systems of the second type ecorii and ecorv. bacteriophage t5 dna is not modified by ecorii and ecorv methylases in vivo. the sites of recognition for restriction endonuclease ecorv are mapped at 24.4; 57.6; 68.5; 70.2% of t5 dna, while the sites at 5.1; 7.6% are recognized by ecorii, the sites at 5.75; 6.0 and 6.5% are recognized by hpai in fst. a high activity of restriction endonucleases ecori and ecorv is demonstrated in crude extracts of e ...	1987	3031490
charons 36 to 40: multi enzyme, high capacity, recombination deficient replacement vectors with polylinkers and polystuffers.	new phage lambda based cloning vectors, charons 36-40, have been constructed which allow cloning of large (up to 24 kb) dna fragments with up to sixteen cloning enzymes. several of these could not be used previously with lambda vectors. clones produced with these vectors can be propagated under recombination deficient conditions. a novel polystuffer method has been developed that permits vector arms to be purified by simple precipitation and which allows reliable identification of clones that ha ...	1987	3031608
cloning and sequence determination of a complementary dna related to human liver microsomal cytochrome p-450 s-mephenytoin 4-hydroxylase.	a cdna sequence related to the human cytochrome p-450 responsible for s-mephenytoin 4-hydroxylation (p-450mp) has been isolated from a human liver bacteriophage lambda gt11 library with antibodies specific for p-450mp. the total length of the cdna is 2.5 kilobases (kb), of which there is a 1.6-kb ecori fragment coding for all but five amino acids corresponding to the n-terminus of the protein and including a small noncoding region at the 3' end. this 1.6-kb fragment has been sequenced and used a ...	1987	3032244
dna-mediated transfer and cloning of a human multidrug-resistant gene of adriamycin-resistant myelogenous leukemia k562.	we have successfully transferred and cloned a fragment of a human multidrug-resistant gene by using dna-mediated gene transfer. macromolecular dna of human multidrug-resistant k562 cells was transfected to drug-sensitive mouse ltk- cells to obtain a drug-resistant transfectant with a human resistant gene. both primary and secondary transfectants showed similar patterns of cross-resistance to adriamycin and vincristine. the mechanism of drug resistance of the transfectants was attributed to decre ...	1987	3032411
correct integration of retroviral dna in vitro.	we have developed a cell-free system for studying the integration of retroviral dna. in our assay, amber mutations in a bacteriophage lambda genome that serves as the target for integration are suppressed by integration of an mlv derivative that carries the e. coli supf gene. the structure of the reaction products is that expected from an authentic mlv integration reaction. linear viral dna from the cytoplasm of infected cells serves as a precursor, though not necessarily the immediate precursor ...	1987	3032450
purification of the cysb protein from salmonella typhimurium.	using an assay based on polypeptide mobility in one- and two-dimensional polyacrylamide electropherograms, we purified the salmonella typhimurium cysb protein from an escherichia coli strain carrying plasmid pgbk14, in which the s. typhimurium cysb gene is under transcriptional control of the strong promoter pl from phage lambda. cysb protein constitutes approximately 4% of total soluble protein in such cells and was obtained with good yield at a purity of 85% after precipitation of nucleic acid ...	1987	3032953
map position and nucleotide sequence of the gene for the large structural phosphoprotein of human cytomegalovirus.	human cytomegalovirus particles contain a phosphoprotein of 150,000 (pp150) apparent molecular weight in their matrix; the protein appears particularly reactive in western blot analyses with human antisera. the gene for pp150 was mapped by screening a bacteriophage lambda gt11 cdna expression library with monospecific rabbit antisera. subsequent hybridization of cdna with cosmid and plasmid clones containing the human cytomegalovirus strain ad169 genome mapped the gene to hindiii fragments j and ...	1987	3033266
cdna clone encoding a high molecular weight antigen of babesia bovis.	an expression library was constructed by inserting cdna copied from mrna of the blood stages of babesia bovis isolate ka into bacteriophage lambda gt11-amp3. an antigen-positive cdna clone detected by screening the library with antibodies from cattle vaccinated with the ka isolate was shown to encode part of a high-molecular weight polypeptide antigen of b. bovis. this molecule was a dominant immunogen and was found by immunofluorescence to be within the parasite in infected erythrocytes.	1987	3033495
toxoplasma gondii and hammondia hammondi: dna comparison using cloned rrna gene probes.	a mung bean nuclease genomic library of purified dna from tachyzoites of the rh strain of toxoplasma gondii was prepared in the bacteriophage lambda gtll and recombinants containing rrna gene fragments were detected by hybridization with radiolabeled total rna from the closely related coccidian eimeria acervulina. ten recombinants were chosen at random, and five of these were investigated further using probes for the genes of the large and small rrna of plasmodium berghei. an insert (called tg4) ...	1987	3034655
transposition studies of mini-mu plasmids constructed from the chemically synthesized ends of bacteriophage mu.	we describe below the chemical synthesis of the right and left ends of bacteriophage mu and characterize the activity of these synthetic ends in mini-mu transposition. mini-mu plasmids were constructed which carry the synthetic mu ends together with the mu a and b genes under control of the bacteriophage lambda pl promoter. derepression of pl leads to a high frequency of mini-mu transposition (5.6 x 10(-2) which is dependent on the presence of the mu ends and the mu a and b proteins. five deleti ...	1986	3034727
the production of generalized transducing phage by bacteriophage lambda.	generalized transduction has for about 30 years been a major tool in the genetic manipulation of bacterial chromosomes. however, throughout that time little progress has been made in understanding how generalized transducing particles are produced. the experiments presented in this paper use phage lambda to assess some of the factors that affect that process. the results of those experiments indicate: the production of generalized transducing particles by bacteriophage lambda is inhibited by the ...	1986	3034738
structural analysis of the locus containing the human c-reactive protein gene and its related pseudogene.	the gene for human c-reactive protein (crp) is mapped within a 34-kilobase pair genomic dna segment identified by chromosome walking through overlapping dna fragments cloned into a lambda phage library. within 16 kilobase pairs upstream and downstream of the locus for the authentic crp gene, only one other sequence homologous to that for crp could be found. sequencing analysis indicates this sequence to be a pseudogene with 50-80% region-specific homology. comparison of the authentic crp gene cl ...	1987	3034876
isolation of the yeast gene encoding elongation factor 3 for protein synthesis.	the gene yef-3 encoding the elongation factor 3 (ef-3) for peptide chain elongation in saccharomyces cerevisiae has been isolated by immunoscreening of a yeast genomic library in the phage lambda gt11. the identity of the ef-3 gene was confirmed by several methods. first, a clone-encoded protein could affinity purify the antibody that specifically reacted with ef-3. second, a recombinant fusion protein, which reacted with anti-beta-galactosidase antibody as well as with anti-ef-3 antibody, was f ...	1987	3034906
regulation of dnab function in dna replication in escherichia coli by dnac and lambda p gene products.	the dnab protein of escherichia coli, a multifunctional dna-dependent ribonucleotide triphosphatase and datpase, cross-links to atp on ultraviolet irradiation under conditions that support rntpase and datpase activities of dnab protein. the covalent cross-linking to atp is specifically inhibited by ribonucleotides and datp. tryptic peptide mapping demonstrates that atp cross-links to only the 33-kda tryptic fragment (fragment ii) of dnab protein. the presence of single-stranded dna alters the co ...	1987	3034907
phage t1-mediated transduction of a plasmid containing the t1 pac site.	the t1 pac site has been cloned into a plasmid vector. this recombinant plasmid was tested for t1-mediated transduction efficiency in comparison with a plasmid containing the phage lambda t1-pac-like site esp-lambda, plasmids containing t1 sequences other than the pac site, and plasmids containing neither t1 sequences nor known pac sites. the data obtained indicate that there are at least two distinct mechanisms of t1-mediated plasmid transduction. one requires the presence of any t1 sequence on ...	1986	3035190
cloning and characterization of the immunity region of phage phi 80.	the immunity region of phage phi 80 has been localized. it codes for at least three proteins: a protein of 34 kda which has the biological properties of the phage repressor, and two other proteins of 9 kda and 18 kda which are the first proteins on the rightward operon. these two proteins are negatively regulated by the 34 kda protein at a divergent promoter site. by position analogy with phage lambda, but not by its biological activity, the 9 kda protein could be the cro product. the 18 kda pro ...	1987	3035344
molecular cloning of cdnas of human liver and placenta nadh-cytochrome b5 reductase.	a cdna coding for human liver nadh-cytochrome b5 reductase (cytochrome b5 reductase, ec 1.6.2.2) was cloned from a human liver cdna library constructed in phage lambda gt11. the library was screened by using an affinity-purified rabbit antibody against nadh-cytochrome b5 reductase of human erythrocytes. a cdna about 1.3 kilobase pairs long was isolated. by using the cdna as a probe, another cdna (pb5r141) of 1817 base pairs was isolated that hybridized with a synthetic oligonucleotide encoding p ...	1987	3035541
electrophoretic mobility of lambda phage hind iii and hae iii dna fragments in agarose gels: a detailed study.		1987	3036265
sequential nmr assignments of labile protons in dna using two-dimensional nuclear-overhauser-enhancement spectroscopy with three jump-and-return pulse sequences.	two-dimensional nuclear overhauser enhancement (noesy) spectra of labile protons were recorded in h2o solutions of a protein and of a dna duplex, using a modification of the standard noesy experiment with all three 90 degree pulses replaced by jump-and-return sequences. for the protein as well as the dna fragment the strategically important spectral regions could be recorded with good sensitivity and free of artifacts. using this procedure, sequence-specific assignments were obtained for the imi ...	1987	3036520
gene 18 protein of bacteriophage t7. overproduction, purification, and characterization.	genetic and physical analyses indicate that gene 18 protein of bacteriophage t7 is essential for packaging of t7 dna. t7 dna is replicated via linear intermediates, culminating in the formation of concatemers many genomes in length which are then packaged into capsids. in infections with phage carrying amber mutations in gene 18, development is blocked at the concatemer stage. biochemical studies on the role of gene 18 protein in concatemer processing and dna packaging have been hampered by its ...	1987	3036832
cloning and mapping of infa, the gene for protein synthesis initiation factor if1.	the gene for translation initiation factor if1, infa, has been identified by using two synthetic oligonucleotides to screen a charon 30 library of escherichia coli dna. a recombinant lambda phage, c1921, was purified from a plaque positive for both probes. a 2.8 kb bglii fragment and a 2.0 kb hindiii fragment isolated from c1921 were subcloned into the bamhi and hindiii sites of pbr322 to yield ptb7 and pth2 respectively. synthesis of if1 in maxicells transformed with ptb7 or pth2 indicates the ...	1987	3037488
isolation of cdna clones coding for human tissue factor: primary structure of the protein and cdna.	tissue factor is a membrane-bound procoagulant protein that activates the extrinsic pathway of blood coagulation in the presence of factor vii and calcium. lambda phage containing the tissue factor gene were isolated from a human placental cdna library. the amino acid sequence deduced from the nucleotide sequence of the cdnas indicates that tissue factor is synthesized as a higher molecular weight precursor with a leader sequence of 32 amino acids, while the mature protein is a single polypeptid ...	1987	3037536
molecular cloning of infectious proviral genomes of bovine leukemia virus.	covalently closed circular dna molecules of bovine leukemia virus were cloned in the lambda phage vector lambda gtwes-lambda b and subsequently in the plasmid vector puc12. proviral dnas of 8.3 kb which have one copy of a long terminal repeat and uniform restriction endonuclease sites were preferentially obtained. four randomly selected clones were examined for their biological activities by dna transfection experiments. the ovine embryonic cells transfected with these clones formed syncytia whi ...	1987	3037775
[a novel oncogene from a human hepatocellular carcinoma].	primary human hepatocellular carcinomas (hcc) were surveyed for oncogenes by transformation assays using nih3t3 cells and the calcium phosphate coprecipitation method. high-molecular-weight dna obtained from the hcc tissues was employed for this purpose. one new transforming dna, named lca (for liver cancer) was obtained, and its properties were studied in detail. this transforming dna had a linkage to the alu sequence and was cloned in lambda phage. restriction enzyme analyses showed that the m ...	1987	3038034
the physical map of the whole e. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library.	thirty-four hundred lambda phage clones containing segments of the e. coli chromosome were isolated and used to construct a 4700 kb long integrated restriction map for eight six-base-recognizing enzymes by a rapid mass-analysis method. our strategy was to measure the sizes of partial restriction enzyme digests by hybridization with a vector probe in a manner analogous to nucleotide sequencing. the data were sorted into groups by a computer program and the boundary clones were further correlated ...	1987	3038334
cloning in escherichia coli of the gene specifying the dna-entry nuclease of bacillus subtilis.	with the aim of cloning genes involved in transformation of bacillus subtilis, a set of transformation-deficient mutants was isolated by means of insertional mutagenesis with plasmid phv60 (vosman et al., 1986). analysis of these mutants showed that those mapping in the aroi region lacked the dna-entry nuclease activity. plasmid phv60 derivatives, containing flanking chromosomal dna fragments, were isolated from these mutants and were used to screen a library of b. subtilis chromosomal dna in ph ...	1987	3038682
a family of lambda phage cdna cloning vectors, lambda swaj, allowing the amplification of rna sequences.	this paper describes the construction and characterization of a family of lambda phage cdna cloning vectors that allows high-efficiency directional cdna cloning and selective amplification of either sense or antisense crna sequences. these vectors contain several unique restriction sites (ecori, xbai, and saci) positioned between two specific phage promoters, sp6 and t7. this system facilitates the in vitro preparation of single-stranded (ss) rna molecules that should be useful in subtractive hy ...	1987	3038683
production of chimeric protein coded by the fused viral h-ras and human n-ras genes in escherichia coli.	a new method for the production of a chimeric protein of two related genes has been developed. the nucleotide sequences of the region from the n terminus to the 86th amino acid (aa) residue of human n-ras and of the harvey sarcoma virus (ha-musv) h-ras are 80% homologous. we isolated the dna fragment encoding the n-terminal portion up to the 70th aa residue from plasmid ph-1 which encodes the total genome of ha-musv, and the dna fragment encoding the c-terminal portion from the 40th aa to the c ...	1987	3038685
inducible expression vectors incorporating the escherichia coli atpe translational initiation region.	new expression vectors were constructed for use in strains of escherichia coli. their most important feature is a polylinker system that facilitates the insertion of a gene in an optimal relationship to the highly efficient e. coli atpe translational initiation region (from nucleotide -50 to the start codon). three atg-containing restriction endonuclease sites can be used for the insertion of the 5' end of a gene at, or near to, its translational initiation codon. these sites may alternatively b ...	1987	3038690
lorist6, a cosmid vector with bamhi, noti, scai and hindiii cloning sites and altered neomycin phosphotransferase gene expression.	four new features have been included in a phage-lambda-replicon-based cosmid vector, lorist6. a scai restriction site allows insertion of blunt-ended dna, partially digested with alui or any other restriction enzyme which creates blunt ends after cleavage. noti and bamhi sites enable construction of libraries using double digestion with noti + sau3ai. the promoter of the neo gene has been substituted by a promoter derived from the tet gene of pbr322, increasing expression of the resistance gene ...	1987	3038694
sequence elements essential for rho-dependent transcription termination at lambda tr1.	to determine the location of dna sequences required for the utilization of rho factor in transcription termination at the tr1 terminator of phage lambda, we constructed and analyzed a series of deletion mutants affecting several distinct regions of the cro-gene sequence. two distinct sequence blocks, ruta and rutb, are shown to be particularly important for rho action at tr1 during in vitro transcription. they are located in the region between the sequence encoding the translation termination co ...	1987	3038914
use of site-specific recombination as a probe of dna structure and metabolism in vivo.	we used site-specific recombination catalyzed by the bacteriophage lambda int system to probe dna structure and metabolism in vivo. in vitro, the complexity of catenated products was linearly proportional to substrate supercoil density. a system was developed that gave efficient, controlled int recombination in escherichia coli cells. from a comparison of the data obtained in vitro and in vivo, we conclude that int recombination does have the same mechanism in vivo as it has in vitro, but that o ...	1987	3039150
specificity of bacteriophage mu excision.	to study the excision of bacteriophage mu at the dna sequence level, the mu-derived phage lambda placmu3 was transposed to the transcribed but non-translated leader region of a plasmid-borne tetracycline (tet) resistance gene. revertants (excision products) were then selected by tet+ restoration of tet+ and characterized. of 21 independent tet+ revertants, 17 contained simple deletions of most or all of lambda placmu3, while the other four contained more complex rearrangements in which one end o ...	1987	3039296
an antitermination protein engages the elongating transcription apparatus at a promoter-proximal recognition site.	as a transcriptional activator, the n protein of phage lambda acts to suppress transcription termination by recognizing a promoter-proximal site, nut, which is separated from the terminators by thousands of base pairs. we demonstrate here that n interacts with the elongating rna polymerase in transit through the boxb domain of nut. this interaction leads to the stable association of n as an integral component of the transcription apparatus. during subsequent elongation, n translocates along with ...	1987	3040263
[transfer of "artificial transposons" constructed on the basis of insertion element is1].	terminal inverted repeats of the insertion element is1 were synthesized chemically and plasmids containing these sequences flanking kanamycin-resistance gene in different combinations were constructed. further incorporation of a whole-sized copy of the is1 into such plasmids caused in some cases the autonomous transfer of km-resistance from plasmid to bacteriophage lambda dna. the transposition of the km-resistance gene was only observed in those cases when the gene was enclosed between is1 copy ...	1987	3040525
expression of a major bovine rotavirus neutralisation antigen (vp7c) in escherichia coli.	sequences from genomic rna segment 8 of the united kingdom tissue-culture (t.c.)-adapted bovine rotavirus encoding a major viral neutralisation antigen vp7c have been expressed in escherichia coli. expression under the regulated control of the bacteriophage lambda pr promoter was as a c-terminal extension to e. coli beta-galactosidase (beta gal). following temperature induction, high levels of the fusion protein were synthesised and accumulated in induced cells, making up 5%-15% of total bacteri ...	1987	3040532
molecular cloning and characterization of the glucosyltransferase c gene (gtfc) from streptococcus mutans lm7.	a glucosyltransferase (gtf) gene, designated gtfc, was cloned from streptococcus mutans lm7. its gene product was detected by screening a bacteriophage lambda library with rabbit antiserum raised against s. mutans lm7 extracellular proteins. dna isolated from the immunopositive recombinant phage revealed two s. mutans chromosomal ecori fragment inserts, 8.1 and 4.7 kilobase pairs in size. escherichia coli minicell analyses revealed the approximate position and direction of transcription of the g ...	1987	3040591
sequence determination and comparison of the exfoliative toxin a and toxin b genes from staphylococcus aureus.	the dna encoding the exfoliative toxin a gene (eta) of staphylococcus aureus was cloned into bacteriophage lambda gt11 and subsequently into plasmid pli50 on a 1,391-base-pair dna fragment of the chromosome. exfoliative toxin a is expressed in the escherichia coli genetic background, is similar in length to the toxin purified from culture medium, and is biologically active in an animal assay. the nucleotide sequence of the dna fragment containing the gene was determined. the protein deduced from ...	1987	3040666
identification of polypeptides encoded by an escherichia coli locus (hfla) that governs the lysis-lysogeny decision of bacteriophage lambda.	we report the cloning of the escherichia coli hfla locus, which governs stability of phage lambda cii protein and which has been proposed to encode or regulate a cii-specific protease. the hfla locus was cloned on an 18-kilobase dna fragment by selecting for plasmids that carry the neighboring pura gene. the boundaries of hfla were delimited by analysis of deletions and insertions constructed in vitro and by use of transposon tn1000. maxicell analysis of the proteins encoded by the hfla-containi ...	1987	3040675
cloning of the gene for the surface array protein of aeromonas salmonicida and evidence linking loss of expression with genetic deletion.	a gene bank of dna from the fish pathogenic bacterium aeromonas salmonicida was constructed in the bacteriophage lambda gt11. phage lambda gt11/10g, a recombinant carrying a 4.0-kilobase fragment of a. salmonicida dna, was found to express the surface array protein (a protein) in escherichia coli. sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the protein expressed from the cloned gene had a subunit molecular weight of 49,000, which was identical to that of subunits in the ...	1987	3040676
characterization of a bacteriophage that carries the genes for production of shiga-like toxin 1 in escherichia coli.	the shiga-like toxin 1-converting bacteriophage h-19b was recently shown to carry the structural genes for the toxin and was shown to have dna sequence homology with phage lambda. we present evidence that the linear genome of bacteriophage h-19b has cohesive termini which become covalently associated during prophage integration. integration occurs through a site on a 4-kilobase-pair ecori fragment located near the center of the bacteriophage chromosome. the relationship between bacteriophages h- ...	1987	3040688
molecular cloning of a 2',3'-cyclic nucleotide 3'-phosphodiesterase: mrnas with different 5' ends encode the same set of proteins in nervous and lymphoid tissues.	antibodies raised to a mixture of the 46 and 48 kda rat cns 2',3'-cyclic nucleotide 3-phosphodiesterases (cnps) recognized apparently identical proteins in peripheral nervous system (pns), thymus, and circulating blood lymphocytes. these antibodies were used to identify, in a rat brain phage lambda gt11 expression library, cdna clones encoding beta-galactosidase-cnp fusion proteins, some of which showed cnp activity. in rna blots, the subcloned cnp cdna inserts hybridized to mrnas of approximate ...	1987	3040924
interactions between escherichia coli rna polymerase and lambda repressor. mutations in prm affect repression of pr.	the rightward operator, or, of bacteriophage lambda is part of a complex regulatory region that includes prm, the promoter for repressor synthesis by a prophage, the rightward early promoter pr, and three repressor-binding sites, or1, or2 and or3. by binding to or2, repressor blocks transcription from pr and simultaneously stimulates the formation of open complexes between rna polymerase and prm. in this letter, we describe a test of the hypothesis that the interaction between rna polymerase bou ...	1988	3045326
cloning and expression of the imipenem-hydrolyzing beta-lactamase operon from pseudomonas maltophilia in escherichia coli.	the l-1 penicillinase structural gene, blas, from pseudomonas maltophilia has been cloned into the vector pacyc184. the pmon01 recombinant plasmid selected by ampicillin resistance carried a 2.6-kilobase sau3a fragment of p. maltophilia dna and was confirmed to express l-1 beta-lactamase by comparative isoelectric focusing. a detailed physical map was constructed, and the blas structural gene was localized with a 17-mer oligonucleotide mixed probe encoding the l-1 n-terminal amino acid sequence. ...	1988	3046482
mutations within the decoding site of escherichia coli 16s rrna: growth rate impairment, lethality and intragenic suppression.	several c----u transitions and small deletions were introduced into the conserved region centered on base c1400 in escherichia coli 16s rrna by in vitro mutagenesis. the mutations were placed within rrnb operons on multicopy plasmids under the transcriptional regulation of either the normal rrnb p1p2 promoters or the temperature-inducible pl promoter from bacteriophage lambda and introduced into e. coli hosts. when expressed from the p1p2 promoters, several of the mutant 16s rrnas impaired cell ...	1988	3047677
aspects of the regulation of adenylate cyclase synthesis in escherichia coli k12.	in escherichia coli k12 expression of the adenylate cyclase gene is subject to multiple controls. in order to gain understanding of the regulation of adenylate cyclase synthesis, operon and protein fusions were constructed by in vitro recombination either into bacteriophage lambda or low-copy-number plasmids, or directly on the chromosome at the cya locus. the fusions were used in physiological experiments as probes to study transcriptional and translational controls of cya expression. it was fo ...	1988	3049935
[effectiveness of expression of chloramphenicol acetyltransferase gene controlled by foreign regulatory regions in escherichia coli. iii. relative effectiveness of cloned promoters of escherichia coli and coliphages].	the study on the rate of initiation of model gene cat transcription under the control of e. coli (plac uv5, ptrp, pcat, ptac), phage lambda (pl, pr), phi x174 (pd) promotors was carried out by means of hybridization of pulse labelled in vivo mrna with the dna coding parts. the presence of gene bla(apr) with its own constitutive promoter in all the recombinants permitted to use the level of appropriate mrna in the cell as an internal standard. this method allowed to evaluate the true efficiency o ...	1988	3054501
deletion and hypermethylation of thymidine kinase gene in v79 chinese hamster cells resistant to bromodeoxyuridine.	previous studies on v79 chinese hamster cells have shown that bromodeoxyuridine (brdu) -resistant variants deficient in thymidine kinase (tk) activity arise by a multistep process which is initiated by a random event and progresses gradually during serial culture in the presence of the drug. in order to determine the molecular basis for the loss of tk activity in these cells, the tk gene was isolated from a lambda phage library of genomic v79 dna, using a fragment of the human tk gene as a probe ...	1988	3057652
in situ detection of sequence-specific dna binding activity specified by a recombinant bacteriophage.	we have used a recombinant bacteriophage that expresses the dna-binding domain of c/ebp to optimize conditions for a screening technique that may facilitate the cloning of genes that encode sequence-specific dna-binding proteins. the method relies on the expression of cdna inserts in bacteriophage lambda gt11. fusion protein adsorbed onto nitrocellulose filters is probed with radioactive, double-stranded dna as a ligand. two procedures greatly increase the level of binding between ligand and rec ...	1988	3061875
tandem gene amplification in vitro for rapid and efficient expression in animal cells.	plasmid vectors phsg293 and phsg747, suitable for in vitro gene amplification for subsequent animal-cell expression, were developed. a cosmid vector phsg293 confers km resistance to escherichia coli host cells and g418 resistance to animal cells and contains a single bstxi recognition/cleavage site, ccacgggg/ctgg, near the cos site (the recognition site is underlined). the cassette vector plasmid phsg747 contains a multiple cloning site (mcs) between the simian virus 40 early promoter and the po ...	1988	3063615
cloning of the bamhi methyl transferase gene from bacillus amyloliquefaciens.	we wish to report the initial characterization of a recombinant clone containing the bamhi methylase gene. genomic chromosomal dna purified from bacillus amyloliquefaciens was partially cleaved with hindiii, fractionated by size, and cloned into psp64. plasmid dna from this library was challenged with bamhi endonuclease and transformed into escherichia coli hb101. a recombinant plasmid pbamm6.5 and a subclone pbamm2.5 were shown to contain the bamhi methylase gene based on three independent obse ...	1988	3063644
development of a trpe promoter-strength measuring system and its use in comparison of the trpedcba, trpr and aroh promoters.	an expression system was developed for measuring in vivo promoter strength at the single copy level and this system was used to compare the trp, aroh and trpr promoters. this system employs trpe enzyme activity as a measure of promoter strength and lacz expression for internal copy number reference. promoter-containing fragments are inserted into a cloning vector and subsequently recombined on to phage lambda by genetic exchange. single lysogens are then prepared and used in promoter-strength an ...	1988	3063826
isolation of the gene for cytochrome p450l1a1 (lanosterol 14 alpha-demethylase) from candida albicans.	the saccharomyces cerevisiae cytochrome p450 l1a1 (lanosterol 14 alpha-demethylase)-coding gene was used as a hybridization probe to isolate two hindiii fragments of 2.5 kb and 6.85 kb from a phage lambda library of candida albicans nucleotide sequences. restriction endonuclease mapping and southern blot hybridization experiments indicated that these fragments represent two allelic forms of the same gene. this cloned sequence, when introduced into s. cerevisiae or c. albicans on a multiple copy ...	1988	3065144
isolation of a phytoalexin-detoxification gene from the plant pathogenic fungus nectria haematococca by detecting its expression in aspergillus nidulans.	detoxification of the pea phytoalexin pisatin via demethylation, mediated by a cytochrome p-450 monooxygenase, is thought to be important for pathogenicity of the fungus nectria haematococca on pea. to isolate a fungal gene encoding pisatin demethylating activity (pda), we transformed aspergillus nidulans with a genomic library of n. haematococca dna constructed in a cosmid which carried the a. nidulans trpc gene. transformants were selected for trp+ and then screened for pda. one transformant a ...	1988	3065148