【摘要】 用PCR技术特异扩增了柔嫩艾美耳球虫TA4抗原基因cDNA序列,克隆至质粒pMD18-T中,获得重组质粒pMD18-T-TA4,采用KpnⅠ/SacⅠ双酶切法及PCR确认正确后,将TA4基因cDNA目的片段亚克隆到乳酸乳球菌表达载体pMG36n中,电穿孔法转化乳酸乳球菌LM0230,获得重组质粒pMG36n-TA4,采用KpnⅠ/SacⅠ双酶切法及DNA测序证明cDNA序列完全正确。获得TA4乳酸乳球菌表达株,经SDS-PAGE电泳分析,结果表明表达产物与预期大小的TA4蛋白分子量一致。更多还原

【Abstract】 TA4 antigen cDNA of E.tenella was amplified by PCR and cloned into a vector pMD18-T.The positive plasmid contained TA4 antigen cDNA was determinded by restriction enzyme analysis and PCR.The plasmid pMD18-T-TA4 and the vector pMG36n were both digested by Sac I and Kpn I.The cDNA encoding TA4 antigen was subcloned into pMG36n and transferred into Lactococcus lactis LM0230 by electroporation.It proved that the positive recombinant plasmid pMG36n-TA4 contained cDNA encoding TA4 antigen by restriction enzyme analysis and sequence determination.In the positive transformants,the TA4 antigen was expressed as a fusion protein in Lactococcus lactis LM0230,which was verified by SDS-PAGE.更多还原