Phasmids as effective and simple tools for construction and analysis of gene libraries

Yankovsky, N. K.; MYu, Fonstein; SYu, Lashina; Bukanov, N O; Yakubovich, Nikita; Ermakova, L. M.; Rebentish, B. A.; Janulaitis, A.; Дебабов, В. Г.

doi:10.1016/0378-1119(89)90180-7

Cited by 13 publications

(6 citation statements)

References 17 publications

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“…A nearly complete (99n5%) EcoRI-generated gene library of M. flagellatus was constructed in the phasmid vector λpSL5 (Yankovsky et al, 1989) in E. coli LE392. The λpSL5 vector with 6-25 kb DNA inserts may exist either as a plasmid when the E. coli host is grown at 30 mC or as phage particles when temperature is shifted to 42 mC.…”

Section: Methodsmentioning

confidence: 99%

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Organization of threonine biosynthesis genes from the obligate methylotroph Methylobacillus flagellatus The GenBank accession numbers for the aspC-hom-thrC-thyA gene cluster and thrB gene sequences from M. flagellatus reported in this paper are L78665 and L78666, respectively.

et al. 1999

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The genes encoding aspartate kinase (ask), homoserine dehydrogenase (hom), homoserine kinase (thrB) and threonine synthase (thrC) from the obligate methylotroph Methylobacillus flagellatus were cloned. In maxicells hom and thrC directed synthesis of 51 and 48 kDa polypeptides, respectively. The hom, thrB and thrC genes and adjacent DNA areas were sequenced. Of the threonine biosynthesis genes, only hom and thrC were tightly linked in the order hom-thrC. The gene for thymidylate synthase (thyA) followed thrC and the gene for aspartate aminotransferase (aspC) preceded hom. All four genes (aspC-hom-thrC-thyA) were transcribed in the same direction. mRNA analysis indicated that hom-thrC are apparently transcribed in one 7 5 kb transcript in M. flagellatus. Promoter analysis showed the presence of a functional promoter between aspC and hom. No functional promoter was found to be associated with the DNA stretch between hom and thrC. The thrB gene encoded an unusual type of homoserine kinase and was not linked to other threonine biosynthesis genes.

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Section: Methodsmentioning

confidence: 99%

“…The λpSL5 vector with 6-25 kb DNA inserts may exist either as a plasmid when the E. coli host is grown at 30 mC or as phage particles when temperature is shifted to 42 mC. Plasmids from the λpSL5 library were transfected into E. coli strains with lesions in threonine biosynthesis genes as described by Yankovsky et al (1989).…”

Section: Methodsmentioning

confidence: 99%

Organization of threonine biosynthesis genes from the obligate methylotroph Methylobacillus flagellatus The GenBank accession numbers for the aspC-hom-thrC-thyA gene cluster and thrB gene sequences from M. flagellatus reported in this paper are L78665 and L78666, respectively.

et al. 1999

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“…Escherichia coli strain DH5~ was used to propagate plasmids pUC18, pUC19, and pBluescriptSK-. The E. coli strain LE392 was used to progagate the phasmid 2pMYF 131 [ 31 ]. Strains carrying the plasmids were grown in SOC medium [24].…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

“…A phasmid library of Synechocystis 6714 DNA prepared according to the method of Yankovsky et al [31] was kindly provided by Dr Michael Fonstein. The phage were screened with the Synechocystis apcAB probe made by the PCR, described above.…”

Section: Library Construction and Sequence Analysismentioning

confidence: 99%

Isolation and characterization of the genes encoding allophycocyanin subunits and two linker proteins from Synechocystis 6714

DiMagno

Haselkorn

1993

Plant Mol Biol

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Genes encoding the phycobilisome core subunits allophycocyanin alpha and beta and a small core linker protein in Synechocystis sp. strain PCC 6714 were cloned and sequenced. These genes form an operon, apcABC, with a single transcription start site and two possible termination sites, one following apcB and the other following apcC. The promoter region, like those of the apcABC operons of other cyanobacteria, does not resemble the consensus promoter sequences of Escherichia coli. However, the apcABC promoters identified in four strains of cyanobacteria have conserved sequences centered at -50 and -10 with respect to the start of transcription. The apcE gene, encoding the protein that links the phycobilisome core to the thylakoid membrane, was also cloned from Synechocystis 6714 and sequenced. It is unlinked to the apcABC operon. As in other Synechocystis strains, the LCM polypeptide encoded by the apcE gene contains three repeats of the basic phycobiliprotein linker domain. The apcE gene promoter sequence bears little resemblance to either the E. coli consensus or the apcABC promoter region, but it is similar to the corresponding regions of other cyanobacterial apcE genes. In these cases, there are conserved sequences centered at -40 and -10 with respect to the transcription start site. These conserved promoter elements from the apcABC and apcE genes were also identified in the corresponding 5'-flanking regions of eleven transcript starts for cpc genes encoding phycocyanin subunits in cyanobacteria and algal chloroplasts. These results suggest that a factor yet to be described participates in transcription of phycobiliprotein genes.

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“…By using cosmids, plasmids containing bacteriophage DNA (Collins and Hohn, 1978;Da vison et ai, 1987;Yankovsky et al, 1989), which can maintain very large DNA inserts, it is now possible to clone the entire genome of complex organisms in a single population of bacteria. By using cosmids, plasmids containing bacteriophage DNA (Collins and Hohn, 1978;Da vison et ai, 1987;Yankovsky et al, 1989), which can maintain very large DNA inserts, it is now possible to clone the entire genome of complex organisms in a single population of bacteria.…”

Section: Research and Developmentmentioning

confidence: 99%

Recombinant plasmids

Hussey¹

1992

Safety in Industrial Microbiology and Biotechnology

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Phasmids as effective and simple tools for construction and analysis of gene libraries

Cited by 13 publications

References 17 publications

Organization of threonine biosynthesis genes from the obligate methylotroph Methylobacillus flagellatus The GenBank accession numbers for the aspC-hom-thrC-thyA gene cluster and thrB gene sequences from M. flagellatus reported in this paper are L78665 and L78666, respectively.

Organization of threonine biosynthesis genes from the obligate methylotroph Methylobacillus flagellatus The GenBank accession numbers for the aspC-hom-thrC-thyA gene cluster and thrB gene sequences from M. flagellatus reported in this paper are L78665 and L78666, respectively.

Isolation and characterization of the genes encoding allophycocyanin subunits and two linker proteins from Synechocystis 6714

Recombinant plasmids

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