O型口蹄疫病毒VP0和VP1蛋白的可溶性表達與反應(yīng)原性分析
發(fā)布時間:2018-06-25 00:10
本文選題:口蹄疫病毒 + VP、VP結(jié)構(gòu)蛋白。 參考:《河南農(nóng)業(yè)科學(xué)》2017年02期
【摘要】:為了高效可溶性表達O型口蹄疫病毒(FMDV)VP0、VP1結(jié)構(gòu)蛋白,根據(jù)大腸桿菌密碼子的偏愛性優(yōu)化合成VP0和VP1基因片段,并將其克隆到p E-SUMO載體中,構(gòu)建重組質(zhì)粒SUMOVP0和SUMO-VP1,將重組質(zhì)粒轉(zhuǎn)化到大腸桿菌BL21(DE3)感受態(tài)細胞中進行誘導(dǎo)表達,并優(yōu)化誘導(dǎo)溫度、時間和IPTG濃度等表達條件。結(jié)果顯示,SUMO-VP0可溶性蛋白表達的最佳條件為:20℃條件下,0.1 mmol/L IPTG誘導(dǎo)表達8 h;SUMO-VP1可溶性蛋白表達的最佳條件為:37℃條件下,0.1 mmol/L IPTG誘導(dǎo)表達12 h。SDS-PAGE電泳和Western blot結(jié)果表明,表達的SUMOVP0、SUMO-VP1可溶性蛋白能夠被抗FMDV的陽性血清識別,具有很好的反應(yīng)原性。
[Abstract]:In order to express VP0 VP1 structural protein of foot-and-mouth disease virus type O (FMDV), VP0 and VP1 gene fragments were synthesized according to the preference of E. coli codon and cloned into pE-SUMO vector. The recombinant plasmids SUMOVP0 and SUMO-VP1 were constructed. The recombinant plasmids were transformed into E. coli BL21 (DE3) cells for induction and expression, and the expression conditions such as induction temperature, time and IPTG concentration were optimized. The results showed that the optimal conditions for the expression of soluble protein of SUMO-VP0 were 0.1 mmol / L IPTG at 20 鈩,
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