Doherty A J, Connolly B A, Worrall A F
Department of Biochemistry, University of Southampton, UK.
Gene. 1993 Dec 22;136(1-2):337-40. doi: 10.1016/0378-1119(93)90491-k.
A synthetic gene coding for bovine pancreatic DNaseI has been cloned under the control of a T7 promoter present on the plasmid pET11. This construct yields a stable Escherichia coli transformant only when transcription from this promoter is tightly controlled. Production of recombinant DNaseI (reDNaseI) is achieved by infection of the cells with a mutant lambda phage, CE6, which carries the gene encoding T7 RNA polymerase. Induced bacterial cultures yield in excess of 2 mg per litre of reDNaseI after purification.
一个编码牛胰腺DNA酶I的合成基因已在质粒pET11上存在的T7启动子的控制下被克隆。只有当该启动子的转录受到严格控制时，这种构建体才能产生稳定的大肠杆菌转化体。通过用携带编码T7 RNA聚合酶基因的突变λ噬菌体CE6感染细胞来实现重组DNA酶I（reDNaseI）的生产。诱导的细菌培养物在纯化后每升产生超过2毫克的reDNaseI。
