泰勒焦虫表面抗原Tams1、SPAG1和TaSP基因的克隆与表达  

Cloning and Expression of Tams1,SPAG1 and TaSP Genes of Theileria annulata Surface Antigen

作　　者：魏玉荣[1] 易忠[1] 马文戈[1] 米晓云[1] 陈智 魏婕[1] 苗书魁[1] 薛英[1] 王延[1] 黄炯[1] 

机构地区：[1]新疆畜牧科学院兽医研究所,乌鲁木齐830000 [2]乌鲁木齐县动物卫生监督所,乌鲁木齐830037

出　　处：《新疆农业科学》2013年第11期2136-2142,共7页Xinjiang Agricultural Sciences

基　　金：新疆维吾尔自治区自然基金青年基金项目(2011211B38);国家自然科学基金项目(31160505)

摘　　要：【目的】以真核表达系统串联表达环形泰勒焦虫表面抗原Tams1、SPAG1和TaSP的高免疫原性区片段,为研究焦虫重组串联蛋白免疫原性奠定基础。【方法】以悬浮培养泰勒焦虫细胞为材料,利用SOEingPCR方法将Tams1、SPAG1和TaSP蛋白高免疫原性区基因通过柔性氨基酸串联嵌合在乙型肝炎病毒核心抗原(HBc)的主要免疫优势区,构建模拟表位-HBc嵌合蛋白真核表达质粒,转染BHK-21细胞,通过Western-Blot检测目的基因表达产物。【结果】Western-Blot分析显示,阳性质粒转染BHK-21细胞裂解产物在硝酸纤维素膜上呈现特异性条带,大小与预期分子量相当;正常BHK-21细胞裂解产物未呈现相应的条带。【结论】重组嵌合蛋白在真核表达系统中成功表达。[ Objective] High immunogenic fragment genes of Tamsl, SPAGI and TaSP were expressed in eukaryotic expression system, which will lay the foundations for recombinant protein immunogenicity of Theile- ria annulata. [ Method ] The genes of high immunogenic fragment genes of Tamsl, SPAG1 and TaSP were am- plified by the SOEing- PCR method using piroplasmosis cells in the suspension culture that was cloned into the major immunodominant region （MIR） of the hepatitis B virus core antigen （HBc） to construct expression plasmids of HBe ehimerism protein. BHK- 21were transfeeted expression plasmids and the product was ana- lyzed by Western -Blot. [ Result]The result of Western -Blot showed that specific bands on the nitrocellulose membrane was present by lysis product of transfected cell. [ Conclusion] It is concluded that chimerism protein was expressed in eukaryotic cell infected with recombinant plasmids successfully.

关 键 词：泰勒焦虫 表面抗原 真核表达 

分 类 号：S852.6[农业科学—基础兽医学]