[Cloning and expression in E.coli of integrase domain of mobile genetic element MDG4 (gypsy)].
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Biomedical subjects
Publications and source records attributed to I L Glukhov.
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The expression of oLpLN region of the plasmid pNT6 causes the high instability of the plasmid. Mutations in the promoter pL region and lesions in the structural part of the N gene result in the stable inheritance of the plasmid. The plasmids pNT6::IS1 containing the IS1-element inserted into the different loci of oLpLN region restore the high instability of the plasmid inheritance in the strain 4830 coding for oLpLN. The plasmids pIG3 and pIG4 of the series pNT6: :IS1 permit one to obtain the collection of random deletions in the cloned fragments induced by IS1-element.
The plasmid pNT6 permits selection of IS1-element insertions into the plasmid occurring with the frequency about 10%. The bacteriophage promoter pL cloned in pNT6 is a hot-spot region for IS1-element insertion. The frequency of IS1 transposition into pL depends on genotype. The plasmid pNT6 may be considered to be a useful target DNA for screening and analysis of IS1-element transposition.
The primary structure of bacteriophage lambda DNA has been searched for the presence of consensus CAP binding sites. Four putative CAP binding sites have been found on the lambda genome, indicating that the catabolite gene activation system of E. coli may be directly involved in the regulation of lambda development. Molecular mechanisms of putative cAMP-CAP-mediated stimulation of lysogenic and lytic responses are discussed.
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A variety of approaches to determine the complete sequence of large DNA fragments is summarized. Main attention is paid to a transposon-based DNA sequencing strategy.
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Cloning vèctor lambda gt-p MB9 has been used for cloning of DNA fragments of bacteriophage T5 produced by EcoR*I activity. One clone contains a DNA fragment of 2.2 Md which has been mapped at 67-71% on the physical map of the genome. Functional studies have shown that bacteriophage lambda gt-T5 can grow on E. coli lights 7. Infection of this E.coli strain with phage lambda gt-T5 induces DNA-ligase activity which has been previously observed in E. Coli infected with bacteriophage T5.