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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

DNA fragments of phage PM2 restricted with Hin dIII endonuclease was cloned in the vector pBR 322 in an Escherichia coli K12 host. The attempt to clone full length PM2 DNA restricted with PstI endonuclease has been unsuccesful. From six randomly chosen recombinant clones DNA was purified and analysed with EcoRI, PstI and Hin dIII endonucleases. The physical map of three chimeric plasmids was unequivocally established. It was shown, that the whole PM2 genome was cloned, although in separate fragments. However, most of the recombinant clones were instable in the absence of selective pressure.
Mol Gen Genet 1979 Nov
PMID:Cloning of bacteriophage PM2 DNA in Escherichia coli K12. 39 43

Plasmids carrying various portions of colicin E1 plasmid (ColE1) DNA have been isolated in an attempt to determine the regions of ColE1 DNA which are required for maintenance of the plasmid in bacteria. To construct the plasmids, the DNA of a ColE1 derivative that contains a gene which controls ampicillin resistance was cleaved by the restriction endonuclease HaeII. The digestion products were joined by T4 DNA ligase and then used to transform bacteria to ampicillin resistance. The plasmid derivatives obtained in this way were always composed of certain HaeII segments. These contain approximately 10% of the ColE1 genome and include the origin of replication of ColE1. We presume that the region of ColE1 which is common to all these derivatives is required for maintenance of the plasmid. After a description of these results, the nucleotide sequence of this region is presented, and possible roles of the region in plasmid replication and maintenance are discussed.
Mol Gen Genet 1979 Oct 03
PMID:Nucleotide sequence of the region required for maintenance of colicin E1 plasmid. 39 52

Comparative analysis of UV-sensitivity was carried out on plasmids of various molecular weight. Recombinant plasmids containing fragments of prokaryotic DNA (E. coli, phage lambda) are repaired in E. coli cells more effectively than those containing eukaryotic DNA fragments. It was also shown that UV-sensitivity of recombinant plasmids is independent of their molecular weight provided that the active repair process in fact occurs. UV-sensitivity was strongly proporational to DNA size only when E. coli double mutant strain was used as host. The survival of plasmids strongly increased after M. luteus UV-endonuclease treatment when E. coli mutant strain uvr- was used as host, but remained practically constant in case of wild type strain. Agarose gel electrophoresis data provide evidence that DNA double-stranded breaks appear un UV-irradiated as well as in UV-endonuclease treated plasmids. One can suggest that UV-inactivation of plasmids results from DNA breaking as a consequence of repair gaps overlap when wild type strain is used as host. Mathematical analysis was carried out assuming this possibility. Experimental data are shown to fit theoretical values calculated assuming that the repair gap size in one DNA strand is equal to 2000-3000 bases.
Mol Biol (Mosk)
PMID:[Repair of recombinant plasmids]. 39

A composite plasmid (pAT2010) has been constructed in vitro from RSF2124 and Bacillus subtilis IFO3022 plasmid (pAT1060) by covalent joining of the two DNA molecules by means of Escherichia coli DNA ligase through the cohesive ends generated by restriction endonuclease RI (EcoRI) cleavage. The composite plasmid was selected by transformation of E. coliC600r-m- with the ligated mixture after enrichment for composite plasmid by preparative agarose gel electrophoresis, and plating of the transformants on a medium containing ampicillin and colicin E1. Treatment of the composite plasmid with EcoRI yielded two fragments corresponding to the linear forms of the parental plasmids. The composite plasmids replicated as biologically functionally units in E. coli, and expressed genetic information carried by RSF2124. In the presence of chloramphenicol, the composite plasmids continued to replicate and the copy number gradually increased. Such nature of replication in the presence of chloramphenicol is characteristic to RSF2124 derived from colicin E1 factor, and so it is suggested that the replicator of RSF2124 is functional in the composite plasmid. The composite plasmid was found to synthesize mRNA of B. subtilis plasmid in cell-free extracts of E. Coli, by hybridization of the mRNA to the original plasmid DNA of pAT1060.
Mol Gen Genet 1977 Nov 29
PMID:Molecular cloning and in vitro transcription of Bacillus subtilis plasmid in Escherichia coli. 41 72

The leucine genes of Bacillus subtilis have been cloned directly from the chromosomal DNA into Escherichia coli leuB cells by selection for the Leu+ phenotype using RSF2124 as a vector plasmid. The hybrid plasmid designated RSF2124-B.leu contained a 4.2 megadalton fragment derived from B. subtilis DNA, including the leu genes. The fragment had one site susceptible to EcoRI* and another site susceptible to BamNI endonuclease. Among the three fragments produced by EcoRI* and BamNI endonucleases, the 1.2 megadalton fragment had the ability to transform B. subtilis leuA, leuB and leuC auxotrophs to leu+. However, B. subtilis ilvB and ilvc auxotrophs were not rescued even by the whole 4.2 megadalton fragment present in the hybrid plasmid. beta-Isopropylmalate dehydrogenase (leuB gene product) activity found in E. coli cells containing the hybrid plasmid was about 60% of that in E. coli wild type cells, despite the high copy number (7.8) of the plasmid per chromosome observed.
Mol Gen Genet 1978 Jan 17
PMID:Cloning of Bacillus subtilis leucina A, B and C genes with Escherichia coli plasmids and expression of the leuC gene in E. coli. 41 24

A study was made of biological and physico-chemical properties of phages of Bac. thuringiensis as well as of a number of parameters of nucleic acids isolated from these phages. The phages contain double-stranded DNA. Molecular weights of DNA from three phages--Tg9, Tg10 and Tg13 have been determined by two independent methods: by measuring the contour length of DNA, from the sedimentation constant and for DNA of phage Tg10 also by endonuclease EcRI hydrolysis. These methods gave similar results. On the basis of the temperature of DNA melting the content of GC pairs was found equal to 37.9, 33.4 and 35.1 mole% for DNA's of phage Tg9, Tg10 and Tg13, respectively. On the basis of measuring the intervals of DNA melting a conclusion was made that DNA of the Tg9 and Tg13 phage has a random distribution of base pairs, while DNA of phage Tg10 displays some clustering of base distribution along the molecule. It has been shown that restrictase EcoRI hydrolyses phage Tg10 DNA into 6 fragments of different molecular weights; DNA's of Tg9 and Tg13 phages are not hydrolyzed. A possibility of existance of phage Tg10 DNA in linear and ring forms has been established. The characteristics of phage particles have been determined by electron microscopy.
Mol Biol (Mosk)
PMID:[Physico-chemical properties of several phages of Bacillus thuringiensis]. 61 32

A study of physico-chemical properties of the nucleic acid of phage Brevibacterium flavum was made. It was found that the phage nucleic acid is a double-stranded DNA. The content of GC pairs was determined by the temperature of DNA melting and by the chromatographic method. In both cases it was (52 +/- 2) mole %. It has been shown that endonuclease Eco RI treatment of phiB DNA forms 12 fragments. The molecular weight of phiB DNA has been determined by two independent methods: by measuring the contous length of DNA and by the sum of molecular weights of DNA restricts obtained after hydrolysis by endonuclease Eco RI and was found to be (30 +/- 1) . 10(6) dalton. Electron microscopy investigations detected rod and circular DNA in either monomeric or dimeric forms. The polarity of DNA arrangement in the phiB phage particle was determined by using the partial denaturation maps.
Mol Biol (Mosk)
PMID:[Physico-chemical study of the nucleic acid of bacteriophage phiB]. 65 78

Upon a 50% isopropanol treatment of phage fl in a 1 M NaCl solution protein A (gene 3 product)--DNA complex is precipitated while protein B (gene 8 product) was still solubilized. After such a treatment the DNA--protein complex containing 10--40% of protein A and less than 0.0025% of protein B was obtained. Evidence was obtained that there was no non-specific rearrangement of protein A during the isolation procedure. The complex was treated with endonuclease R.HAC III, followed by electrophoresis of the resulted fragments and estimation of the [14C] protein A (labeled with [14C]histidine) throughout the gel. The maximal radioactivity coincided with the DNA bands, being proportional to the DNA content in the respective bands. The data obtained indicate that protein A is iniformly arranged along the DNA molecule.
Mol Biol (Mosk)
PMID:[Complex of single-stranded phage DNA and protein--a product of bacteriophage fl gene 3. Distribution of gene 3 protein in the DNA molecule]. 66 19

Plasmid inter-relationships were studied by hybridisation of a radioactively labelled DNA probe to endonuclease-derived fragmentation patterns of plasmids bound to a nitrocellulose filter. The degradative plasmids SAL and NAH were found to be very closely related, but probably one did not give rise to the other by just a single deletion or insertion. Relationships between SAL and other degradative plasmids are complex; substantial homology was found with TOL and other plasmids encoding toluate dissimilation and significant homology was found with OCT.
Mol Gen Genet 1978 Apr 17
PMID:Molecular relationships of degradative plasmids determined by in situ hybridisation of their endonuclease-generated fragments. 67 96

DNA fragments generated by the EcoRI of HindIII endonucleases from the low copy number antibiotic resistance plasmids R6 and R6-5 were separately cloned using the high copy number ColE1 or pML21 plasmid vectors and the insertional inactivation procedure. The hybrid plasmids that were obtained were used to determine the location of the EcoRI and HindIII cleavage sites on the parent plasmid genomes by means of electron microscope heteroduplex analysis and agarose gel electrophoresis. Ultracentrifugation of the cloned fragments in caesium chloride gradients localized the high buoyant density regions of R6-5 to fragments that carry the genes for resistance to streptomycin-spectinomycin, sulfonamide, and mercury and a low buoyant density region to fragments that carry the tetracycline resistance determinant. Functional analysis of hybrid plasmids localized a number of plasmid properties such as resistances to antibiotics and mercury and several replication functions to specific regions of the R6-5 genome. Precise localisation of the genes for resistance to chloramphenicol, kanamycin, fusidic acid and tetracycline was possible due to the presence of identified restriction endonuclease cleavage sites within these determinants. Only one region competent for autonomous replication was identified on the R6-5 plasmid genome and this was localized to EcoRI fragment 2 and HindIII fragment 1. However, two additional regions of replication activity designated RepB and RepC, themselves incapable of autonomous replication but capable supporting replication of a linked ColE1 plasmid in polA- bacteria, were also identified.
Mol Gen Genet 1978 Jun 14
PMID:Cloning and characterization of EcoRI and HindIII restriction endonuclease-generated fragments of antibiotic resistance plasmids R6-5 and R6. 67


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