鸡importin β1基因真核表达载体的构建及亚细胞定位  

Construction of Eukaryotic Expression Vector and Subcellular Localization of Chicken importin β1 Gene

作　　者：孙鑫 吴燕芝 董霁 冒勖 颜修万 熊建民[1] 段志强[1,2] SUN Xin;WU Yanzhi;DONG Ji;MAO Xu;YAN Xiuwan;XIONG Jianmin;DUAN Zhiqiang(College of Animal Science, Guizhou University, Guiyang, Guizhou 550025;Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang, Guizhou 550025)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州大学高原山地动物遗传育种与繁殖教育部重点实验室,贵州贵阳550025

出　　处：《中国家禽》2018年第4期21-24,共4页China Poultry

基　　金：国家自然科学基金项目(31502074和31760732);大学生"SRT计划"项目(贵大SRT字[2015]177号);贵州大学引进人才科研项目(贵大人基合字[2014]10号)

摘　　要：根据GenBank已发表的鸡importinβ1基因序列,设计合成一对特异性引物从重组克隆质粒pCR2.1-importin β1中扩增鸡importinβ1基因开放阅读框,通过酶切、连接等方法将其亚克隆到真核表达载体pEGFP-C1构建重组真核表达载体pEGFP-importin β1。利用脂质体转染法将pEGFP-importin β1转染DF-1细胞,24h后分别检测重组蛋白EGFP-importin β1在细胞中的表达及亚细胞定位情况。结果表明,成功构建了鸡importin β1基因的重组真核表达载体pEGFP-importin β1;将其转染DF-1细胞后得到与预期大小相符的EGFP-importin β1重组蛋白条带,荧光显微镜观察显示重组蛋白主要定位在细胞核。研究结果为进一步开展鸡importin β1蛋白与相关家禽病毒蛋白的相互作用研究奠定了工作基础。A pair of specific primers was designed to amplify the ORF region of chicken importin β1 gene from the cloning plasmid pCR2.1-importin 151. The obtained PCR products were then subcloned into the eukaryotic expression vector pEGFP-C1 to construct the recombinant plasmid pEGFP-importin β1 by enzyme digestion and ligation methods. The expression and subcellular localization of the recombinant protein EGFP-importin β1 were detected at 24 h after the recombinant plasmid was transfected into DF-1 cells. The results showed that the recombinant plasmid pEGFP-importin β1 was successfully constructed and the expected recombinant protein EGFP-importin β1 was obtained when the recombinant plasmid was transfected into cells. Fluorescence analysis indicated that the recombinant protein EGFP-importin β1 mainly localized in the nucleus. These results laid the foundation for further studying the interaction between chicken importin 151 protein and other viral proteins.

关 键 词：鸡 IMPORTIN Β1基因 真核表达 亚细胞定位 

分 类 号：S831[农业科学—畜牧学] S855.3[农业科学—畜牧兽医]