畜牧兽医学报2011，Vol.42Issue(8)：1071-1080,10.
p21基因在牛卵成熟中的表达检测及其真核表达载体构建
Detection for p21 Gene Expression in Bovine Oocytes Maturation and Construction of Eukaryotic Expression Vector pVenus-P21
摘要
Abstract
The objectives of this study were to determine expression of p21 gene during IVM of bovine oocytes, and construct a eukaryotic expression vector pVenus-P21. Firstly, the RT-PCR and immunohistochemical staining were applied to check the expression of p21 mRNA and protein at different maturation stages of bovine oocytes. Then the p21 gene was cloned from bovine fi-broblasts by RT-PCR, and a eukaryotic expression vector pVenus-P21 was constructed. After pVenus-P21 mediated by Lipofectamine 2000, it was transfected into Hela cells, it was efficiently expressed, which was identified by Fluorescence microscopy observation and RT-PCR. In addi-ation, the cRNA of p21-venus transcripted in vitro was microinjected into bovine oocytes, and the expression and location was observed under fluorescence microscopy. In conclusion, these data demonstrate that p21 mRNA and protein are expressed at defferent stages of bovine oocytes. Besides, the recombinant vector pVenus-P21 is successfully constructed. After transfection or mi-croinjection, the fusion protein could be expressed efficiently and localized accurately both in Hela cells and bovine oocytes. This greatly facilitates the further study of P21, especially its role onoocytes maturation and early embryo development.关键词
牛卵母细胞/p21基因/体外转录/显微注射Key words
bovine oocyte/ p21 gene/ transcription in vitro/ microinjection分类
畜牧业引用本文复制引用
赵贵民,杨文琳,吴苏君,云彦,雷安民..p21基因在牛卵成熟中的表达检测及其真核表达载体构建[J].畜牧兽医学报,2011,42(8):1071-1080,10.基金项目
2009-2011年西北农林科技大学青年科学基金项目(Z109021004) （Z109021004）
