畜牧兽医学报2013，Vol.44Issue(1)：15-22,8.
鸡Piwi蛋白的亚细胞定位研究
Sub-cellular Localization of Chicken Piwi Protein
摘要
Abstract
The localization of Piwi protein in eukaryotic cells was conducted by constructing two eukaryotic expression vectors, aimed to provide basis for further study on the biological functions of Piwi protein. CDS of Piwi gene from Langshan chicken testicular tissue was cloned by Reverse Transcript-PCR. Fusion expression vector containing the enhanced green fluorescent protein (EGFP), pEGFP-C1-Piwi, was constructed; then was transfected into NIH-3T3 with Lipofectin PEL Meanwhile, the eukaryotic expression vector pcDNA3. 1-Piwi was also constructed successfully, and then Piwi protein expression was detected in NIH-3T3 cells by indirect immunofluores-cence assay. CDS of Piwi gene was cloned successfully, and the sequence size was 2 604 bp, which was similar with GenBank. Fusion expression vector pEGFP-C1-Piwi expressed in the whole NIH-3T3 cells, while Piwi protein of pcDNA3. 1-Piwi was found to locate in the cytoplas-mic of NIH-3T3 cells by indirect immunofluorescence assay. Piwi protein mainly expressed in the cytoplasmic of NIH-3T3 cells, and indirect immunofluorescence assay was more reliable than the EGFP reporter gene method, which was fit for investigating sub-cellular localization of target protein.关键词
Piwi蛋白/真核表达/亚细胞定位/绿色荧光蛋白/间接免疫荧光Key words
Piwi protein/ eukaryotic expression/ subcellular localization/ EGFP/ indirect immunofluorescence分类
农业科技引用本文复制引用
戴爱琴,常国斌,陈蓉,张颖,阴彦辉,马腾,李建超,陈国宏..鸡Piwi蛋白的亚细胞定位研究[J].畜牧兽医学报,2013,44(1):15-22,8.基金项目
国家自然科学基金项目(31172199 （）
30972088) （）
江苏省自然科学基金(BK2009190) （BK2009190）
江苏高校优势学科建设工程资助项目 （）
