农业生物技术学报2012，Vol.20Issue(1)：54-61,8.DOI:10.3969/j.issn.1674-7968.2012.01.008
牛体外受精胚胎早期发育差异表达基因的筛选及定量检测
Screening and Quantitative Analysis of Differential Expression Genes in In vitro Fertilized Bovine Pre-implantation Embryos
摘要
Abstract
The large capacity of differential expression genes during oocyte maturation and the transition from maternal to zigotic control makes it possible to screen genes playing crucial roles in embryonic development by mRNA differential display reverse transcription PCR (DDRT-PCR). In order to explore the gene expression mechanism of ire vitro maturation (FVM) of oocytes and in vitro development of pre-implantation embryos derived from in vitro fertilization (FVF) in different developmental stages in cattle, mRNA DDRT-PCR was employed to screen differential expression genes in germinal vesicle (GV) stage oocytes, 8-cell stage embryos and blastocysts. Real-time quantitative PCR (Q-PCR) was used to analyze the expression abundance of the differential expression genes in the three stages and in vitro matured M II oocytes. Five differential displayed straps were discovered by DDRT-PCR. The results of sequencing and database analysis showed that the 5 fragments were corresponding to pyrophosphatase (inorganic) 1 {PPA1), erbb2 interacting protein (ERBB2IP), restin-like (CEP350), ribosomal protein L27a (RPL27A) and IK cytokine, down-regulator of HLAII (IK), respectively. Q-PCR and SPSS statistical analysis indicated that the mRNA level of PPA1, ERBB21P and CEP350 decreased along with the procedure of embryonic development, whereas that of RPL27A turned to increase till blastocyst stage. The mRNA level of IK fluctuated before blastocyst stage and then decreased. Overall, the results of this research provide more information on effects of oocyte maturation on the following embryonic development and the mechanism of zygotic gene activation.关键词
卵母细胞成熟/胚胎发育/差异基因/mRNA差异显示/实时定量PCRKey words
Oocyte maturation/Embryo development/Differential expression genes/ mRNA differential display/ Rea-time Quantitative PCR Q-PCR引用本文复制引用
刘冬,沙日娜,温建勋,王瑞,仓明,刘东军..牛体外受精胚胎早期发育差异表达基因的筛选及定量检测[J].农业生物技术学报,2012,20(1):54-61,8.基金项目
国家基础科学人才培养基金(No.J0730648)、国家自然科学基金(No.30760168)和内蒙古自然科学基金重大项目(No.200711020401) （No.J0730648）
