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Abstract

The gene of methylase M.SccL1I that protects DNA against hydrolysis with the nickase N.BspD6I was inserted into plasmid pRARE carrying genes of tRNA, which are rare in E. coli. The insertion of the gene sscML1I into pRARE was reasoned by incompatibility of pRARE and the plasmid carrying the gene sscML1I, because both plasmids contained the same ori-site. Upon transformation of E. coli TOP10F' cells with both the recombinant plasmid pRARE/MSsc and the expression vector pET28b containing the nickase gene bspD6IN under the phage T7 promoter, a strain of E. coli was obtained which produced 7 ·10^sup 5^ units of the nickase N.BspD6I per 1 g wet biomass, and this yield was two orders of magnitude higher than the yield of the enzyme from the strain free of pRARE/MSsc.[PUBLICATION ABSTRACT]

Details

Title
Plasmid pRARE as a Vector for Cloning to Construct a Superproducer of the Site-Specific Nickase N.BspD6I
Author
Rogulin, E A; Perevyazova, T A; Zheleznaya, L A; Matvienko, N I
Pages
1123-7
Publication year
2004
Publication date
Oct 2004
Publisher
Springer Nature B.V.
ISSN
00062979
e-ISSN
16083040
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
719076092
Copyright
MAIK "Nauka/Interperiodica" 2004