猪流行性腹泻病毒M蛋白的原核表达
发布时间：2018-10-23 09:09
【摘要】：本试验设计了M基因的特异性引物,扩增了M基因,成功构建了重组质粒,并转化至Escherichia coli BL21(DE3)感受态细胞中,优化了异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达条件.结果表明,重组表达的融合蛋白Pet-32a-M大小约为43 ku,与预期大小相符.十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)检测表明,M蛋白为包涵体蛋白,且在IPTG浓度为0.8 mmol·L-1,温度为37℃,诱导时间为8 h的条件下,蛋白表达效果最佳.蛋白免疫印迹检测结果进一步显示,重组蛋白Pet-32a-M在体外可以被成功表达.
[Abstract]:The specific primers of M gene were designed, the M gene was amplified, the recombinant plasmid was successfully constructed and transformed into Escherichia coli BL21 (DE3) competent cells, and the induced expression conditions of isopropyl- 尾 -D-thiogalactoside (IPTG) were optimized. The results showed that the Pet-32a-M size of the recombinant fusion protein was about 43 ku,. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that M protein was the inclusion body protein, and the protein expression was best under the conditions of IPTG concentration 0.8 mmol L -1, temperature 37 鈩，
本文编号：2288776
[Abstract]:The specific primers of M gene were designed, the M gene was amplified, the recombinant plasmid was successfully constructed and transformed into Escherichia coli BL21 (DE3) competent cells, and the induced expression conditions of isopropyl- 尾 -D-thiogalactoside (IPTG) were optimized. The results showed that the Pet-32a-M size of the recombinant fusion protein was about 43 ku,. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that M protein was the inclusion body protein, and the protein expression was best under the conditions of IPTG concentration 0.8 mmol L -1, temperature 37 鈩，
本文编号：2288776
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