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[Cloning and regulation of gene expression of EcoRV restriction- modification system].
Mol Biol (Mosk). 1990 Mar-Apr; 24(2):438-47.MB

Abstract

A number of recombinant plasmids, containing EcoRV restriction-modification genes have been constructed. Individual genes of this system were introduced into plasmids of various incompatibility groups. Promoter regions of genes encoding methylase and restrictase have been cloned and studied. With the use of specialized vector pVE8 it was shown that the efficiency of the endonuclease gene promoter is comparable with early lambda phage promoters and produced about 70% of PL efficiency. The efficiency of the methylase gene promoter region was twice less than the efficiency of the restriction endonuclease gene promoter. Plasmid with restriction endonuclease gene promoter located downstream in relation to the additional regulatable phage lambda promoter PL has been obtained. It enabled us to construct strains 30-40 fold overproducing this enzyme under conditions of inactivation of the temperature sensitive phage repressor c1857. This construction directs the production of a high level (10%) of the total cellular soluble proteins) of the EcoRV restriction enzyme. The factors that influenced the level of enzyme synthesis under induction are discussed.

Authors

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Pub Type(s)

English Abstract
Journal Article

Language

rus

PubMed ID

2194115

Citation

Kravets, A N., et al. "[Cloning and Regulation of Gene Expression of EcoRV Restriction- Modification System]." Molekuliarnaia Biologiia, vol. 24, no. 2, 1990, pp. 438-47.
Kravets AN, Zakharova MV, Solonin AS, et al. [Cloning and regulation of gene expression of EcoRV restriction- modification system]. Mol Biol (Mosk). 1990;24(2):438-47.
Kravets, A. N., Zakharova, M. V., Solonin, A. S., Kuz'min, N. P., Taniashin, V. I., Glatman, L. I., Moroz, A. F., & Baev, A. A. (1990). [Cloning and regulation of gene expression of EcoRV restriction- modification system]. Molekuliarnaia Biologiia, 24(2), 438-47.
Kravets AN, et al. [Cloning and Regulation of Gene Expression of EcoRV Restriction- Modification System]. Mol Biol (Mosk). 1990 Mar-Apr;24(2):438-47. PubMed PMID: 2194115.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Cloning and regulation of gene expression of EcoRV restriction- modification system]. AU - Kravets,A N, AU - Zakharova,M V, AU - Solonin,A S, AU - Kuz'min,N P, AU - Taniashin,V I, AU - Glatman,L I, AU - Moroz,A F, AU - Baev,A A, PY - 1990/3/1/pubmed PY - 1990/3/1/medline PY - 1990/3/1/entrez SP - 438 EP - 47 JF - Molekuliarnaia biologiia JO - Mol. Biol. (Mosk.) VL - 24 IS - 2 N2 - A number of recombinant plasmids, containing EcoRV restriction-modification genes have been constructed. Individual genes of this system were introduced into plasmids of various incompatibility groups. Promoter regions of genes encoding methylase and restrictase have been cloned and studied. With the use of specialized vector pVE8 it was shown that the efficiency of the endonuclease gene promoter is comparable with early lambda phage promoters and produced about 70% of PL efficiency. The efficiency of the methylase gene promoter region was twice less than the efficiency of the restriction endonuclease gene promoter. Plasmid with restriction endonuclease gene promoter located downstream in relation to the additional regulatable phage lambda promoter PL has been obtained. It enabled us to construct strains 30-40 fold overproducing this enzyme under conditions of inactivation of the temperature sensitive phage repressor c1857. This construction directs the production of a high level (10%) of the total cellular soluble proteins) of the EcoRV restriction enzyme. The factors that influenced the level of enzyme synthesis under induction are discussed. SN - 0026-8984 UR - https://www.unboundmedicine.com/medline/citation/2194115/[Cloning_and_regulation_of_gene_expression_of_EcoRV_restriction__modification_system]_ L2 - http://rebase.neb.com/rebase/refs/1247.html DB - PRIME DP - Unbound Medicine ER -