山羊骨骼肌特异性基因α-actin启动子驱动的Fat-1表达载体及其构建方法  

The expression vector of Fat-1 driven by specific α-actin promoter in goat skeletal muscle and construction of the vector

作　　者：孟繁星[1] 樊懿萱[1] 钟部帅[1] 王锋[1] MENG Fanxing;FAN Yixuan;ZHONG Bushuai;WANG Feng(Collega of Animal Science and Technology,National Experimental Teching Demonstration Center for Animal Science,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物科技学院,动物科学类国家级实验教学示范中心,江苏南京210095

出　　处：《畜牧与兽医》2020年第12期1-8,共8页Animal Husbandry & Veterinary Medicine

基　　金：国家肉羊产业技术体系(CARS-38)。

摘　　要：试验旨在构建山羊骨骼肌特异性基因α-actin启动子驱动的Fat-1表达载体。将Fat-1基因克隆入pEGFP-N1载体,构建成为pEGFPN1-Fat-1载体;插入CMV增强子和骨骼肌特异性基因α-actin启动子,构建成为α-actin promoter-pEGFPN1-Fat-1表达载体;最后通过流式细胞仪检测转Fat-1基因对山羊耳成纤维细胞周期和凋亡的影响。双酶切和测序证实,已成功构建了肌肉组织特异性表达载体α-actin promoter-pEGFPN1-Fat-1;与对照组相比,转Fat-1基因促进羊耳成纤维细胞增殖,抑制凋亡。试验结果为动物转基因工程的研究和应用提供了参考依据。This study was to construct a Fat-1 expression vector driven by theα-actin promoter of specific goat skeletal muscle gene.In the experiment,the Fat-1 gene was cloned into the pEGFP-N1 vector to construct a pEGFPN1-Fat-1 vector.Then the CMV enhancer and theα-actin promoter of the specific skeletal muscle gene were inserted to construct anα-actin promoter-pEGFPN1-Fat-1 expression vector.Finally,the effect of transfection of the Fat-1 gene on goat ear fibroblast proliferation and the cells was detected using flow cytometry.The results showed that double enzyme digestion and sequencing confirmed that the muscle tissue-specific expression vectorα-actin promoter-pEGFPN1-Fat-1 had been successfully constructed.Compared with the control group,the transfection of the Fat-1 gene promoted the proliferation of sheep ear fibroblasts and inhibitd apoptosis,which laid a foundation for future applications of the vector in genetic engineering.

关 键 词：骨骼肌 启动子驱动 Fat-1过表达载体 山羊 

分 类 号：S827[农业科学—畜牧学]