新疆多浪羊pcDNA3.1-IL-1β表达载体的构建及鉴定  

Construction and identification of expression vector pcDNA3.1-IL-1β of Duolang sheep in Xinjiang

作　　者：张晶[1] 宋显明[1] 李玮[1] 艾东旭 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]塔里木畜牧科技兵团重点实验室,新疆阿拉尔843300

出　　处：《黑龙江畜牧兽医》2017年第1期102-106,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(30960277);研究生科研创新项目(TDGRI201520);南疆地区羊标准化养殖的群发病防控及预警响应机制研究项目(TDZKCX201401)

摘　　要：为了构建新疆多浪羊真核表达载体pc DNA3.1-IL-1β,试验采用重组质粒p MD-18TIL-1βPCR和质粒pc DNA3.1双酶切的方法,获得目的基因白细胞介素-1β(IL-1β)和载体片段pc DNA3.1(+),通过连接、转化宿主菌大肠杆菌DH5α感受态细胞,挑取阳性克隆,再通过菌液PCR和双酶切来验证阳性克隆,并测序。结果表明:试验已成功构建真核表达载体pc DNA3.1-IL-1β,完成了pc DNA3.1-IL-1β的菌液PCR和酶切鉴定。通过对IL-1β功能的了解,以及熟悉质粒提取和质粒连接的方法,完成重组质粒pc DNA3.1-IL-1β的构建。To construct a eukaryotic expression vector pcDNA3.1 - IL - 1β of Duolang sheep in Xinjiang, target gene interleukin - 1β （ IL - 1β） and vector fragment pcDNA3.1 （ ＋ ） were obtained using recombinant plasmid pMD - 18T - IL - 1β by PCR and plasmid pcDNA3.1 by double digestion method in the test, and then the IL - 1β and pcDNA3.1 （ ＋ ） were ligated and transformed into Escherichia coli DH5α com- petent cells. The positive clones were selected and verified by bacteria liquid PCR of bacteria and double enzyme digestion, and then se- quenced. The results showed that the eukaryofic expression vector pcDNA3.1 - IL - 1β was successfully constructed, and the bacteria liquid PCR and double enzyme digestion of pcDNA3.1 - IL - 1β were completed. The construction of recombinant plasmid pcDNA3.1 - IL - 1β was accomplished by the understanding of IL - 1β function and the method of plasmid extraction and plasmid ligation.

关 键 词：真核表达载体 构建质粒 酶切鉴定 白细胞介素-1 β(IL-1 β) 多浪羊 

分 类 号：Q786[生物学—分子生物学] S826[农业科学—畜牧学]