【摘要】：本試驗設(shè)計了M基因的特異性引物,擴(kuò)增了M基因,成功構(gòu)建了重組質(zhì)粒,并轉(zhuǎn)化至Escherichia coli BL21(DE3)感受態(tài)細(xì)胞中,優(yōu)化了異丙基-β-D-硫代半乳糖苷(IPTG)誘導(dǎo)表達(dá)條件.結(jié)果表明,重組表達(dá)的融合蛋白Pet-32a-M大小約為43 ku,與預(yù)期大小相符.十二烷基硫酸鈉聚丙烯酰胺凝膠電泳(SDS-PAGE)檢測表明,M蛋白為包涵體蛋白,且在IPTG濃度為0.8 mmol·L-1,溫度為37℃,誘導(dǎo)時間為8 h的條件下,蛋白表達(dá)效果最佳.蛋白免疫印跡檢測結(jié)果進(jìn)一步顯示,重組蛋白Pet-32a-M在體外可以被成功表達(dá).
[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，

本文編號：2288776