球形芽孢杆菌R修饰甲基化酶在大肠杆菌中的过量表达及其纯化至均一性。

Overproduction of the Bacillus sphaericus R modification methylase in Escherichia coli and its purification to homogeneity.

作者信息

Posfai G, Kiss A, Venetianer P

出版信息

Gene. 1986;50(1-3):63-7. doi: 10.1016/0378-1119(86)90310-0.

Abstract

A DNA fragment containing the information coding for the GGCC-specific Bacillus sphaericus R modification methylase, BspR, was inserted into plasmid vector pKK223-3 under the control of the strong and inducible tac promoter, and transformed into Escherichia coli HB101. Upon induction this strain accumulated the methylase enzyme (while cell growth was inhibited) up to several percent of total cellular protein. Homogeneous methylase could be prepared in three purification steps.

摘要

将一段包含编码GGCC特异性球形芽孢杆菌R甲基化酶BspR信息的DNA片段，插入到受强诱导型tac启动子控制的质粒载体pKK223 - 3中，并转化到大肠杆菌HB101中。诱导后，该菌株积累甲基化酶（同时细胞生长受到抑制），其含量可达细胞总蛋白的百分之几。通过三个纯化步骤可制备出纯的甲基化酶。