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目的检测重组人B7-H1IgV(rhB7-H1)工程菌生物学特性的稳定性。方法将重组质粒pQE30-B7-H1IgV转化的大肠杆菌rhB7-H1/DH5α工程菌连续传代50代,每隔10代进行工程菌B7-H1IgV蛋白表达量检测、扫描电镜观察、革兰染色、各项生化反应及质粒的酶切鉴定。结果各代工程菌中B7-H1IgV蛋白的表达量均为菌体总蛋白的10%左右;各代工程菌提取的质粒双酶切后均可见381 bp的目的基因片段;各代工程菌均呈典型的大肠杆菌形态,革兰染色显示为阴性杆菌,各项检测结果与原始菌种无显著差异。结论该工程菌生物学特性稳定,可作为生产用菌种用于大规模生产。
Objective To detect the stability of the biological characteristics of recombinant human B7-H1IgV (rhB7-H1) engineered bacteria. Methods The recombinant plasmid pQE30-B7-H1IgV was transformed into E. coli rhB7-H1 / DH5α for 50 generations. The expression of B7-H1IgV protein was detected every 10 passages. The results of scanning electron microscopy, Biochemical reactions and plasmid digestion identification. Results The expression levels of B7-H1IgV protein in all the engineered bacteria were all about 10% of the total bacterial proteins. The 381 bp target gene fragments could be seen after double digestion of the engineered bacteria in each generation. Typical Escherichia coli morphology, Gram stain showed negative bacilli, the test results no significant difference with the original species. Conclusion The biological characteristics of the engineering bacteria are stable and can be used as large-scale production for the production of bacteria.