【作者】 张莉； 秦泽荣； 徐镔蕊； 王春凤； 何召庆； 刘尚高；

【Author】 Zhang Li ,Qin Zerong,Xu Binrui ,He Zhaoqing,Wang Chunfeng,Liu Shanggao (China Agricultural University , Beijing 100094 ,China)

【机构】 北京市林科学院； 中国农业大学动物医学院；

【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.获得的质粒pMD18-T-TA4.采用KpnI/SaeI双酶切法及PCR确认正确后,将TA4基因cDNA目的片段亚克隆到表达戴体pMG36n中,电穿孔法转化乳酸乳球菌LM0230.获得质粒pMG36n-TA4.采用KpnI/SaeI双酶切法及DNA测序证明eDNA序列完全正确。获得TA4乳酸乳球菌表达珠.SDS-PAGE分析表明该菌球表达预期大小的TA4抗原蛋白。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 SacI and KpnI. The cDNA encoding TA4 antigen was subcloned into pMG36n and transferred into Lactococcus lactisLM0230 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 antigea was expressed as fusion protein in Lactococcus lactis LM0230,which was verified by SDS-PAGE.更多还原