论文部分内容阅读
目的 :在大肠杆菌宿主系统中表达 Tropic180 8基因编码的蛋白。方法 :用 PCR方法从质粒中扩增出 Tropic180 8c DNA开放阅读框架片段 (82 8bp) ,并重组入表达载体 p ET-2 1a中 ,转化大肠杆菌 BL2 1(DE3) ,用 IPTG诱导 Tropic180 8融合蛋白的表达 ,经 SDS-PAGE,表达蛋白条带转至 PVDF膜进行氨基酸序列分析。结果 :构建了 p ET-2 1a-180 8重组质粒 ,外源性 Tropic180 8基因可在大肠杆菌中获得表达 ,目的蛋白占细菌总蛋白的 14 %以上 ,表达蛋白的 N端氨基酸序列与基因编码序列一致。结论 :Trop-ic180 8基因可在大肠杆菌中获得表达 ,这为进一步纯化该基因表达蛋白、制备相应抗体以及研究其结构与功能奠定了基础
OBJECTIVE: To express the protein encoded by the Tropic 180 gene in E. coli host system. Methods: The open reading frame fragment of Tropic180 8c DNA (82 8bp) was amplified from the plasmid by PCR and recombined into the expression vector p ET-2 1a. The recombinant plasmid was transformed into E. coli BL21 (DE3) and Tropic 1808 fusion was induced by IPTG After SDS-PAGE, the protein bands were transferred to PVDF membrane for amino acid sequence analysis. Results: The recombinant plasmid pET-2 1a-180 8 was constructed and the exogenous Tropic 180 8 gene was expressed in E. coli. The target protein accounted for more than 14% of total bacterial proteins. The N-terminal amino acid sequence of the expressed protein was The same sequence. CONCLUSION: The Trop-ic180 8 gene can be expressed in E. coli, which lays the foundation for further purification of the expressed protein, preparation of the corresponding antibody and study of its structure and function