如皋黄鸡功能未知基因C1EIP表达载体的构建及其抗小鼠血清制备  

Construction of eukaryotic expression vector for Rugao yellow chicken undefined gene C1EIP and preparation of antiserum

作　　者：李东[1] 王飞[1] 纪艳芹 汪怡临 王曼[1] 张晨[1] 王颖洁[1] 何娜娜[1] 靳锴 路镇宇 程少泽 张亚妮[1] 李碧春[1] 

机构地区：[1]扬州大学动物科学技术学院,江苏扬州225009

出　　处：《中国兽医学报》2017年第6期1161-1166,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31272429 31472087);高等学校博士学科点专项科研基金资助课题资助项目(20103250110006);江苏省"六大人才高峰"资助项目;江苏省优势学科资助项目

摘　　要：为了深入研究鸡胚胎干细胞(embryonic stem cell,ESC)向雄性生殖细胞分化过程,基于RNA-Seq基础,筛选出功能未知且差异极其显著的关键基因C1EIP,PCR克隆得到其编码区全长序列409bp,构建其真核表达载体pcDNA3.0-C1EIP和pEGFP-C1EIP,实现C1EIP在真核细胞内的定位表达,并以PEI包裹pcDNA3.0-C1EIP后免疫小鼠制备抗血清。间接免疫荧光检测抗血清效价为1∶10,RT-PCR和Western blot证实C1EIP真核表达载体成功表达且抗血清效果良好,为C1EIP基因在ESCs向雄性生殖细胞分化过程中的功能研究奠定了基础。Based on RNA-Seq, extremely significant different function-unknown gene, CIEIP was screened,and full-length CDS sequence 409 bp was amplified by polymerase chain reaction（PCR）. Then the eukaryotic expression vector pcDNA3.0-C1EIP and pEGFP-C1EIP were established, lo- cation expression of pEGFP-C1EIP was perform, pcDNA3.0-CIEIP was packaged with PEI and used to immunize mice. Protein CIEIP was expressed as expected and the chicken anti-mouse C1EIP antiserum was obtained. The antiserum titer reaching 1 ： 10 was identified by IFA. Besides, Western blot analysis showed that the antiserum could be used to detect protein C1EIP. This study can contribut to further research on the undefined gene C1EIP.

关 键 词：如皋黄鸡 C1EIP 功能未知基因 真核表达 抗血清 

分 类 号：S814.8[农业科学—畜牧学]