【摘要】 将牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)p20和p14基因分别亚克隆至原核表达载体pGEX-6p-1和pPROEX-HTb,并转化至相应的宿主菌大肠杆菌BL21(DE3)pLysS和DH5α中表达。P20蛋白在2个宿主中都获得了高效表达;P14蛋白在BL21(DE3)pLysS中表达量较低,而在DH5α中未见表达。对P14蛋白诱导表达过程的监测表明,P14蛋白对大肠杆菌是一种毒性蛋白。用Western-blotting分析未能检测到两种蛋白的反应条带,推测这两种蛋白可能存在构象表位。更多还原

【Abstract】 The P20 and P14 protein genes of bovine viral diarrhea virus（BVDV） were subcloned into two prokaryotic expression vector,pGEX-6p-1 and pPROEX-HTb,respectively,to construct recombinant expression plasmids.The recombinant plasmids were transformed into competent BL21（DE3）pLysS and DH5α accordingly for expression.Protein P20 was highly expressed in each of the host bacteria induced by IPTG.P14 protein was expressed at low level in BL21（DE3）pLysS,but not expressed in DH5α.Detection of D_600nm values in the course of expression of the P14 protein showed that the P14 protein was toxic to Escherichia coli.It was proposed that these two proteins may contain conformational epitopes because no reaction was observed using Western-blotting.更多还原