[天蚕素X在大肠杆菌中的融合表达及凝血因子Xa的切割作用]
[Fusion expression of cecropin X including the cleavage of FXa in Escherichia coli].
作者信息
Yuan L D, Dou F, Liang Y P, Xie W, Wang F, Zhang S Q, Dai Z Y
机构信息
Department of Biochemistry, Nanjing University.
出版信息
Sheng Wu Gong Cheng Xue Bao. 2000 May;16(3):411-4.
PCR method was used to introduce the code sequence of Factor Xa cleavage site to the 5' end of cecropin CMIV mutant gene X, then the gene was cloned into the expression vector pGEX-KG, and was highly expressed in E. coli BL21 by IPTG induction. The fusion protein was purified by affinity-chromatography and was cleaved by Factor Xa. Cecropin X with antibacterial activity was obtained after purified by ion-exchange chromatography.
采用聚合酶链反应（PCR）方法将凝血因子Xa切割位点的编码序列引入天蚕素CMIV突变体基因X的5'端，然后将该基因克隆到表达载体pGEX-KG中，并通过异丙基-β-D-硫代半乳糖苷（IPTG）诱导在大肠杆菌BL21中高效表达。融合蛋白通过亲和层析纯化，并用凝血因子Xa进行切割。经离子交换层析纯化后获得具有抗菌活性的天蚕素X。
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