山羊FGF1基因腺病毒超表达载体的构建  

Construction of Adenovirus Vector of FGF1 Gene in Goat

作　　者：黄凯 林亚秋[1] 朱江江 马洁琼 王永[1,2] HUANG Kai;LIN Ya-qiu;ZHU Jiang-jiang;MA Jie-qiong;WANG Yong(Department of Life Science and Technology,Southwest Minzu University,Sichuan Chengdu 610041,China;Protection and Utilization of Animal genetic Resources in Qingzang Plateau Sichuan Provincial Key Lab,Sichuan Chengdu 610041,China)

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041 [2]青藏高原动物遗传资源保护与利用四川省重点实验室,四川成都610041

出　　处：《西南农业学报》2018年第11期2418-2422,共5页Southwest China Journal of Agricultural Sciences

基　　金：四川省"十三五"畜禽育种攻关项目(2016NYZ0045)

摘　　要：【目的】为利用超表达手段进一步研究成纤维细胞生长因子1(FGF1)基因对山羊肌内前体脂肪细胞分化过程的影响。【方法】本试验通过构建p Ad Track-CMV-FGF1穿梭质粒,转入含有腺病毒骨架载体pAdEasy-1的大肠杆菌BJ5183感受态细胞中进行同源重组,获得腺病毒超表达载体pAdEasy-FGF1,经Pac I酶切鉴定后转染293A细胞进行病毒的包装与扩繁,最后用包装的pAdEasy-FGF1感染山羊肌内前体脂肪细胞,检测感染前后FGF1的表达变化。【结果】成功构建了腺病毒超表达载体p AdEasyFGF1,并在293A细胞中成功包装,其感染山羊肌内前体脂肪细胞能显著上调FGF1 mRNA的表达水平。【结论】说明FGF1基因可以通过感染重组腺病毒表达载体pAdEasy-FGF1在山羊肌内前体脂肪细胞中实现超表达。[ Objective ] The present paper was conducted to further study the effect of fibroblast growth factor 1 （ FGF1 ） gene on the differen- tiation of intramuscular preadipocyte in goat by overexpression. [ Method ] In this experiment, pAdTrack-CMV-FGF1 shuttle plasnfid was constructed. The adenovirus superexpression vector pAdEasy-FGF1 was obtained by homologous recombination of Escherichia coli BJ5183 receptive cells containing adenovirus skeleton vector pAdEasy-1. Plasnfid pAdEasy-FGF1 was digested with PacI and then being transfected into 293A cells for virus packaging and propagation. Finally, pAdEasy-FGF1 was used to infect goat intramuscular preadipocytes. [ Result] The adenovirus superexpression vector pAdEasy-FGF1 was successfully constructed and packaged in 293A cells. The infection of goat intra- muscular precursor adipocytes can significantly up-regulate FGF1. [ Conclusion] The expression level of mRNA suggests that FGF1 gene can be overexpressed in goat intramuscular preadipocytes by infection with recombinant adenovirus expression vector pAdEasy-FGF1.

关 键 词：山羊 成纤维细胞生长因子1 晾病毒 

分 类 号：S858.26[农业科学—临床兽医学]