柔嫩艾美耳球虫TA4抗原基因cDNA在乳酸乳球菌中的克隆与表达  

Cloning and expression of TA4 antigen from Eimeria tenella in Lactococcus lactis

作　　者：张莉[1] 秦泽荣[2] 崔尚金[3] 何召庆[2] 刘尚高[2] 

机构地区：[1]北京市农林科学院畜牧兽医研究所,北京100089 [2]中国农业大学动物医学院,北京100094 [3]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2006年第3期271-274,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金资助项目(30070571)

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。TA4 antigen eDNA of E.tenella was amplified by PCR and cloned into a vector pMDIS-T. The positive plasmid contained TA4 antigen eDNA was determinded by restriction enzyme analysis and PCR. The plasmid pMDI8-T-TA4 and the vector pMG36n were both digested by Sac Ⅰ and Kpn Ⅰ, The eDNA 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 eDNA encoding TA4 antigen by restriction enzyme analysis and sequence determination. In the positive transformants, the TA4 antigen was expressed as a fusion protein in Laetoeoecus lactis LM0230, which was verified by SDS-PAGE.

关 键 词：柔嫩艾美耳球虫 TA4抗原基因cDNA 乳酸乳球菌 克隆与表达 

分 类 号：S852.61[农业科学—基础兽医学]