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目的克隆结核分枝杆菌MPT63抗原蛋白的编码基因Rv1926c,并在大肠杆菌中表达纯化,获得结核分枝杆菌重组MPT63蛋白。方法以结核分枝杆菌H37Rv基因组为模板,PCR扩增Rv1926c基因编码序列,克隆入原核表达载体pET-30a,构建重组表达质粒,转化大肠杆菌后用IPTG诱导表达,纯化表达产物,获得重组MPT63抗原蛋白。结果测序表明重组质粒pET30aRv1926c具有正确的编码序列,质粒构建成功,重组蛋白MPT63在大肠杆菌中以包涵体形式稳定表达。结论结核分枝杆菌MPT63重组蛋白能在大肠杆菌工程菌种成功表达,为进一步的应用研究奠定基础。
Objective To clone the gene Rv1926c encoding Mycobacterium tuberculosis MPT63 antigen and express it in E.coli to obtain the recombinant Mycobacterium tuberculosis MPT63 protein. Methods The Mycobacterium tuberculosis H37Rv genome was used as a template to amplify the coding sequence of Rv1926c gene and cloned into the prokaryotic expression vector pET-30a. The recombinant plasmid was constructed and transformed into E.coli. The recombinant plasmid was induced with IPTG and the recombinant protein was purified. The recombinant MPT63 antigen protein. Results The sequencing showed that the recombinant plasmid pET30aRv1926c had the correct coding sequence and the plasmid was successfully constructed. The recombinant protein MPT63 was stably expressed as inclusion body in E. coli. Conclusion The recombinant protein of Mycobacterium tuberculosis MPT63 can be successfully expressed in Escherichia coli engineering strains, which lays the foundation for further application research.