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牛瑟氏泰勒虫p33双拷贝基因的真核表达
引用本文:杨兴,许应天,严昌国,高旭,孙平杰,张丽霞.牛瑟氏泰勒虫p33双拷贝基因的真核表达[J].中国兽医学报,2011,31(7).
作者姓名:杨兴  许应天  严昌国  高旭  孙平杰  张丽霞
作者单位:延边大学农学院,吉林延吉,133002
基金项目:延边大学“211工程”三期重点学科基金资助项目
摘    要:为构建牛瑟氏泰勒虫p33双拷贝基因真核表达质粒,以牛瑟氏秦勒虫基因组DNA为模板,应用SOE—PCR技术得到牛瑟氏泰勒虫p33双拷贝基因,将其先克隆到pMD18T—Simple载体,再亚克隆到真核表达载体pVAX1中,得到重组质粒pVAX1—2p33。对该重组质粒进行PCR、酶切鉴定及测序后,采用脂质体法将其转染到BHK-21细胞,用RTPCR和IFA进行鉴定。结果表明,牛瑟氏泰勒虫p33双拷贝基因真核表达质粒pVAX12p33构建成功并可以在BHK-21细胞中表达。本试验结果为p33双拷贝基因的免疫学研究奠定了基础。

关 键 词:牛瑟氏泰勒虫  p33双拷贝基因  pVAX1—2p33  真核表达

Prokaryotic expression of p33 double-copy genes from Theileria sergenti
YANG Xing,XU Ying-tian,YAN Chang-guo,GAO Xu,SUN Ping-jie,ZHANG Li-xia.Prokaryotic expression of p33 double-copy genes from Theileria sergenti[J].Chinese Journal of Veterinary Science,2011,31(7).
Authors:YANG Xing  XU Ying-tian  YAN Chang-guo  GAO Xu  SUN Ping-jie  ZHANG Li-xia
Institution:YANG Xing,XU Ying-tian,YAN Chang-guo,GAO Xu,SUN Ping-jie,ZHANG Li-xia(Agricultural College,Yanbian University,Yanji,Jilin 133002,China)
Abstract:To construct bovine Theileriosis sergenti p33 double-copy genes eukaryotic expression plasmid,we take bovine Theileriosis sergenti genome DNA as the template and apply SOE-PCR technique to get the bovine Theileriosis sergenti p33 double-copy genes,after that clone the p33 double-copy genes to the pMD18-T-Simple vector,then subclone the eukaryotic expression vector pVAX1,finally get a recombinant plasmid pVAX1-2p33.Apply PCR to the recombinant plasmid and identify by enzyme cutting and sequencing,then transf...
Keywords:Theileria sergenti  p33 double-copy genes  pVAX1-2p33  prokaryotic expression  
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