目的：通过对肺炎链球菌TruB（SpTruB）基因进行原核表达重组质粒的构建，在大肠杆菌中表达SpTruB蛋白，再运用凝胶迁移实验（EMSA）对TruB与tRNA的相互作用进行检测。方法 PCR扩增SpTruB基因序列，然后转入E.coli BL21中进行表达，利用镍离子亲和层析纯化，采用EMSA检测SpTruB与tRNA的相互作用。结果正确构建了SpTruB基因的表达质粒，SpTruB蛋白在E .coli BL21中为可溶性上清表达，成功纯化了目的蛋白，EMSA检测显示SpTruB能与tRNA进行相互作用。结论 SpTruB能与tRNA进行相互作用。%Objective To analysis the codon by constructing the expressing plasmids, and the SpTruB protein was expressed with solution in E .coli system,to detect interactions about tRNA and TruB by EMSA. Methods SpTruB was amplified by PCR, and then expressed by transforming into E .coli BL21,the plasmids were constructed by mutant the rare codes into normal ones. The expressed production was purified by nickel column chromatography. Results The expression vector of the SpTruB was successfully constructed and the expressed aim protein in E .coli BL21 in a soluble supernatant way. EMSA analysis showed that SpTruB could interact with tRNA. Conclusion SpTruB could interact with tRNA.
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