p21 基因在牛卵成熟中表达检测及真核.pdfVIP

2011, 42(8) : Acta Veterinaria et Zootechnica Sinica p 21 赵贵民, 杨文琳, 吴苏君, 云 彦, 雷安民* ( , , , 712100) : p 21 , RTPCR p 21 mRNA p 21 pVenusP21 2000 pVenusP21 H ela , RTPCR , H ela p 21venus cRNA, , , , p 21 mRNA pVenusP21 H ela p 21v e nus cRNA , P21 : ; p 21 ; ; : S823; Q3 . 13 :A : 0366696 ( 2011) Detection forp 21 Gene Expression in Bovine Oocytes Maturation and Construction of Eukaryotic Expression Vector pVenus 21 * ZHAO Guimin, YANG Wenlin, WU Sujun, YUN Yan, LEI Anmin (Shanxi Key Laboratory of Molecular Biology f or Agriculture, Shanxi Stem Cell ngineering and Technology Research Center, Collegeof Veterinary Medicine, N orthwest A F University , Yangling 712100, China) Abstract: T he objectives of this study w ere to determine expression of p 21 gene during IVM of bovine oocytes, and construct a eukaryotic expression vector pVenusP21. Firstly , the RTPCR and immunohistochemical staining w ere applied to check the expression of p 21 mRNA and protein at different maturation stages of bovine oocytes. Then the p 21 gene w as cloned from bovine fi broblasts by RTPCR, and a eukaryotic expression vector pVenusP21 w as constructed. A fter pVenusP21 mediated by Lipofectamine 2000, it w as transfected into Hela cells, it w as efficiently expressed, w hich w as identified by Fluorescence microscopy observation and RTPCR. In addi ation, the cRNA of p 21v enus transcripted inv itro w as microinjected into bovine oocytes, and the expression and location w as observed under fluorescence microscopy . In conclusion, these data demonstrate that p 21 mRNA an