卡拉库尔羊TNF-α基因原核表达载体的构建及表达条件优化  

Construction of Prokaryotic Vector of TNF-α Gene from Karakul Sheep and Optimization of Expression Conditions

作　　者：贺艳艳[1] 潘辉[1] 王娟娟 左文斌[1] 李莲瑞[2,3] 

机构地区：[1]塔里木大学生命科学学院,新疆阿拉尔843300 [2]塔里木大学动物科学学院,新疆阿拉尔843300 [3]新疆生产建设兵团塔里木畜牧科技重点实验室,新疆阿拉尔843300

出　　处：《湖北农业科学》2013年第7期1686-1688,共3页Hubei Agricultural Sciences

基　　金：新疆生产建设兵团应用基础项目(07GC07)

摘　　要：利用反转录PCR(RT-PCR)技术扩增卡拉库尔羊肿瘤坏死因子α(TNF-α)基因,连接到pET-28b载体,转化大肠杆菌BL21(DE3),在不同的IPTG浓度、时间、温度条件下诱导TNF-α的表达。结果表明,原核表达载体pET-28b-TNF-α构建成功,可在大肠杆菌BL21(DE3)中诱导表达,优化的诱导表达条件为诱导温度37℃、诱导时间4 h、IPTG浓度1.0 mmol/L。TNF-α(tumor necrosis factor α) gene of Karakul sheep was cloned by reverse transcription PCR(RT-PCR),linked to pET-28b vector and then used to transform E.coli BL21(DE3).Expression of TNF-α was induced under different IPTG concentration,time and temperature.The results showed that the prokaryotic expression vector pET-28b-TNF-α was successfully constructed;and its express could be induced in E.coli BL21(DE3) cells.The optimized expression conditions were induction temperature,37 ℃;induction time,4 h;and IPTG concentration,1.0 mmol / L.

关 键 词：肿瘤坏死因子α(TNF-α) 原核表达 表达条件 

分 类 号：S[农业科学]