绵羊IL-6启动子的克隆及pIL6-TLR4载体构建  

Cloning of Sheep IL-6 promoter and the Construction of the pIL6-TLR4 Vector

作　　者：宗炎[1] 阚童童 潘求真[1] 王静宜[1] 连正兴[2] Zong Yan;Kan Tongtong;Pan Qiuzhen;Wang Jingyi;Lian Zhengxing(Heilongjiang Bayi Agricultural University,Daqing 163319;College of Animal Science and Technology,China Agricultural University)

机构地区：[1]黑龙江八一农垦大学,大庆163319 [2]中国农业大学动物科技学院

出　　处：《黑龙江八一农垦大学学报》2017年第1期45-48,共4页journal of heilongjiang bayi agricultural university

基　　金：国家重大转基因专项(2014ZX08008-005)

摘　　要：为生产抗病转基因绵羊,并确保转基因羊的安全性。克隆绵羊IL-6启动子序列,以IL-6为启动子构建TLR4过表达载体,转染绵羊成纤维细胞验证功能。用1μg·L-1的LPS刺激转染后的绵羊成纤维细胞,利用q PCR技术和ELISA方法检测受LPS刺激后的成纤维细胞TLR4 m RNA表达量和TLR4蛋白表达量。结果表明,转染后的TLR4 m RNA表达量在2、4、24、48 h显著高于对照组(P<0.05),其中2、4、24 h极显著高于对照组(P<0.01),TLR4蛋白表达量始终高于对照组。说明p IL6-TLR4过表达载体构建成功。To produce disease resistance transgenic sheep and to ensure its security,the sheep IL-6 promoter sequences were cloned.The TLR4 over -expression vector was constructed by using sheep IL -6 as a promoter,and transfected into sheep fibroblasts successfully.The transfected sheep fibroblasts was stimulated with 1 μg·L-1 of LPS,and the technology of qPCR was used to detect the mRNA and protein expression. The results showed that,the mRNA expressions of TLR4 in transfected sheep fibroblasts after 2,4,24,48 h were significantly higher than that in the control group(P<0.05),in which after 2,4,24 h were extremely significantly higher(P<0.01),meanwhile the TLR4 protein expression was always higher than that in the control group. It was proved that the pIL6-TLR4 over-expression vector was constructed successfully.

关 键 词：TLR4 IL-6启动子 转基因 绵羊 

分 类 号：Q78[生物学—分子生物学]