腺病毒介导人卵泡刺激素基因在山羊乳腺上皮细胞的表达及蛋白活性分析  

Protein Activity Analysis and Expression of Human Follicle Stimulating Hormone Factor in Goat Mammary Epithelial Cells by Adenovirus

作　　者：李媛[1] 华荣茂 段家昕 陈华丽[1] 徐德军 杨丽 程建勇[1] 李晓亚 耿果霞[2] 李青旺[1] LI Yuan;HUA Rongmao;DUAN Jiaxin;CHEN Huali;XU Dejun;YANG Li;CHENG Jianyong;LI Xiaoya;GENG Guoxia;LI Qingwang(College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,China;College of Veterinary Medicine,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《家畜生态学报》2020年第10期18-23,共6页Journal of Domestic Animal Ecology

基　　金：转基因生物新品种培育重大专项(2018ZX0801013B)。

摘　　要：该研究旨在构建含人FSH基因的重组腺病毒载体,在山羊乳腺上皮细胞中进行表达,验证重组人FSH的体外生物学活性。利用AdEasy重组腺病毒系统获得pAdEasy-FSH/His重组腺病毒载体,经HEK293A细胞包装、扩增后获得重组人FSH腺病毒,感染GMEC后,检测FSH表达情况。结果表明,将获得的较高滴度的含FSH目的基因的重组腺病毒感染GMEC后,经Western blot检测GMEC细胞可以成功表达FSH蛋白;通过体外细胞活性试验验证其具备生物学活性。综上所述,腺病毒介导的羊乳腺上皮细胞表达的重组人FSH具备体外生物学活性,这为进一步在大动物乳腺表达重组FSH奠定了基础。The aim of this study was to construct a recombinant adenoviral vector containing the human FSH gene,which was expressed in goat mammary epithelial cells.And the biological activity of recombinant human FSH was verified in vitro.The pAdEasy-FSH/His recombinant adenoviral vector was obtained using the AdEasy recombinant adenovirus system.The recombinant adenoviral vector was packaged and amplified by HEK293A cells to obtain recombinant human FSH adenovirus.After the obtained recombinant human FSH adenovirus infected GMEC,the expression of FSH was detected.The results showed that the recombinant adenovirus containing the higher titer FSH gene was infected with GMEC,and the FSH protein was successfully expressed by Western blot.It had been verified to have biological activity by cell viability assay in vitro.In summary,recombinant human FSH expressed by adenovirus-mediated goat mammary epithelial cells had biological activity in vitro.This would lay a foundation for further expression of recombinant FSH in large animal mammary glands.

关 键 词：FSH 腺病毒表达系统 羊乳腺上皮细胞 生物活性 

分 类 号：S811.5[农业科学—畜牧学]