OSP-1启动子在奶山羊卵巢颗粒细胞中的转录活性  

Transcriptional activity of ovarian-specific promoter from rat in dairy goat granulosa cells

作　　者：白龙[1] 刘海艳[1] 赵海波[1] 彭甲银[1] 曹斌云[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《中国兽医学报》2013年第4期630-635,共6页Chinese Journal of Veterinary Science

基　　金：国家科技支撑计划资助项目(2011BAD28B05-3)

摘　　要：采用大鼠OSP-1启动子替换掉pcDNA3.1(+)中的CMV启动子,在其下游插入EGFP基因片段,构建载体pOSP-1-EGFP,脂质体转染不同细胞,利用荧光检测、RT-PCR以及相对定量PCR鉴定该启动子启动基因表达的活性。结果表明,在卵巢颗粒细胞、人卵巢癌细胞系HO-8910中可看到有绿色荧光;OSP-1片段只在卵巢颗粒细胞和人卵巢癌细胞系HO-8910有表达,并在人卵巢癌细胞系HO-8910中表达量较高,但是两者之间差异不显著。说明OSP-1启动子可调控外源基因在奶山羊卵巢颗粒细胞中特异性的表达。Taking pcDNA3, 1（＋） as skeleton, OSP-1 promoter from rat was employed to replace CMV promoter for the construction of the reporter vector pOSP-1-EGFP,in which EGFP was used as a reporter to examine the activity of OSP-1 in GCs. EGFP was successfully expressed in dairy goat GCs and HO-8910 cell line transfected with pOSP-1-EGFP. Meanwhile, RT-PCR and Real- time PCR analysis further proved the tissue specific transcription of EGFP mRNA in those two cell types. However,the difference of EGFP expression in goat GCs and HO-8910 cell line was not significant. It is concluded that OSP-1 promoter from rat can specifically drive foreign genes expression in diary goat GCs. Thus, we obtained a tissue specific regulation element and provided a potential tool for research of regulation and development of ovary in dairy goat.

关 键 词：OSP-1 EGFP 卵巢颗粒细胞 RT-PCR 

分 类 号：S827[农业科学—畜牧学]