Overexpression of a lethal methylase, M.TneDI, in E. coli BL21(DE3)
作者： Hui Xu , Dongmei Han , Zhaohui Xu
DOI: 10.1007/S10529-014-1552-Z
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摘要: A pET-based vector pDH21 expressing the methylase, M.TneDI (recognizing CGCG) from Thermotoga was constructed, and transformed into E. coli BL21(DE3). Despite BL21(DE3) being McrBC positive, 30 transformants were isolated, which suspected to be McrBC− mutants. The overexpression of verified by SDS-PAGE analysis. Compared previously constructed pJC340 vector, a pACYC184 derivative tet promotor, newly improved expression methylase about fourfold, allowing complete protection DNA substrates. This study not only demonstrates practical approach overexpressing potential lethal proteins in but also delivers production strain that may useful various vitro methylation applications.
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