人泛素基因原核表达载体构建、表达及鉴定

目的]构建含人泛素(Ubiquitin,Ub)基因的原核表达载体,并在大肠杆菌表达系统中表达。方法]应用PCR扩增人Ub基因片段,插入表达载体pGEX-4T-1中,转化大肠杆菌BL21(DE3)PLysS,经诱导表达及鉴定。结果]PCR扩增出约228bp的基因片段,克隆至载体后,经测序与GenBank中的一致,表达蛋白相对分子质量约34000,纯化后鉴定为目的蛋白。结论]已成功获得人Ub蛋白,为进一步的研究和应用奠定了基础。

Construction, Expression and Characterization on Prokaryotic Expression Vector of Human Ubiquitin Gene

Objective] The aim of the paper was To construct the prokaryotic expression vector of human ubiquitin gene and express the gene in E.coli.Method] The Ub gene was Amplified by PCR, then was inserted into expression vector pGEX-4T-1 to transform to E.coli BL21(DE3) for expression. Result] A gene fragment at a length of 228 bp was amplified. And its sequence was identical to that reported in GenBank. The protein with a relative molecular weight of 34 000 was expressed and was identified as Ub after purification. Conclusion] Ub protein was successfully expressed. It laid a foundation for further study and application of superantigen Ub.