Panasenko S M, Cameron J R, Davis R W, Lehman I R
Science. 1977 Apr 8;196(4286):188-9. doi: 10.1126/science.322281.
A lambda vector that contains the gene for Escherichia coli DNA ligase (lambdagt4-lop-11 lig+) has been modified to achieve overproduction of this enzyme. The third Eco RI site in the lambda chromosome has been altered by mutation, and the left-hand Eco RI fragment has been shortened. The new vector, lambdagt4-lop-11 lig+, forms a stable lysogen which, upon induction, produces a 100-fold increase in DNA ligase activity. Introduction of a phage mutation (S7) that prevents cell lysis results in an even greater increase (500-fold).
一个含有大肠杆菌DNA连接酶基因(λgt4 - lop - 11 lig +)的λ噬菌体载体已被改造,以实现该酶的过量表达。λ噬菌体染色体中的第三个Eco RI位点已通过突变改变,并且左侧的Eco RI片段已被缩短。新载体λgt4 - lop - 11 lig +形成稳定的溶原菌,诱导后,DNA连接酶活性增加100倍。引入一个阻止细胞裂解的噬菌体突变(S7)会导致更大幅度的增加(500倍)。