鸡Stra8基因真核表达载体构建及亚细胞定位的研究  

Construction of Eukaryotic Expression Vector and Sub-cellular Location of Stra8 Gene in Chicken

作　　者：王丹[1] 张振韬[1] 施青青[1] 黄晓梅[1] 朱才业[1] 郑蒙蒙[1] 李伟[1] 徐剑蓉 王克华[3] 李碧春[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009 [2]扬州市邗江区畜牧兽医站,江苏扬州225002 [3]中国农业科学院家禽研究所,江苏扬州225125

出　　处：《中国家禽》2013年第8期10-13,共4页China Poultry

基　　金：国家自然科学基金(31272429)

摘　　要：试验旨在克隆苏禽黄鸡视黄酸激活基因8(Stra8)的cDNA序列,构建pEGFP-C1-Stra8载体并制备鸡Stra8抗体。通过RT-PCR克隆苏禽黄鸡Stra8基因cDNA,构建pEGFP-C1-Stra8和pcDNA3.1(+)-Stra8重组质粒,分别转染NIH-3T3细胞和免疫小鼠制备抗体,间接免疫荧光法检测抗体效价以及在细胞中的分布。结果显示,苏禽黄鸡Stra8基因cDNA全长645bp,共编码214个氨基酸,提交至GenBank获得登录号为JX204292.1;成功制备Stra8多克隆抗体,pEGFP-C1-Stra8融合蛋白在NIH-3T3细胞质中表达。该结果为深入探讨禽类Stra8生物学功能奠定了基础。The aim of the study was to clone cDNA of Stra8 in Suqin yellow chicken,construct pEGFP-C1-Stra8 and prepare polyelonal antibody against Stra8. The cDNA of Stra8 gene of Suqin yellow chicken was cloned by RT-PCR, and the recombinant eukaryotic expression vectors pEGFP-C1-Stra8 and pcDNA3.1 （＋） -Stra8 were constructed and used for NIH-3T3 cells transfection and mouse immune,respectively. Serum antibody titer and subcellular localization were detected by indirect immunofluorescent assay. The eDNA of Stra8 had a length of 645 bp encoding 214 amino acids. The sequence was submitted to GenBank with accession number JX204292.1. The polyclonal antibody against Stra8 had been prepared successfully; the conjugated protein was efficiently expressed in the cytoplasm of NIH-3T3 cells. These results had a good value for further research of Stra8 function.

关 键 词：Stra8基因 质粒DNA免疫 多克隆抗体 亚细胞定位 

分 类 号：S572[农业科学—烟草工业]