人脂联素基因启动子荧光素酶报告基因载体的构建  

Cloning of Human Adiponectin Promoter and Construction of Luciferase Reporter Vectors

作　　者：路玲玲[1,2] 宰军华[1] 崔琳[1] 李强[3] 张莎莎[1] 金小琴[2] 

机构地区：[1]河南中医学院第一附属医院,郑州450000 [2]河南中医学院第一临床医学院,郑州450003 [3]河南中医学院第一附属医院/河南省病毒性疾病中医药防治重点实验室,郑州450000

出　　处：《世界科学技术-中医药现代化》2013年第1期55-61,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：郑州市科技公关计划项目(0910SGYS33391-1):以脂肪细胞分泌因子为靶点的减肥中药初步筛选;负责人:宰军华

摘　　要：目的:克隆不同长度人脂联素基因(AD)启动子片段,构建含人AD启动子片段的荧光素酶报告基因载体并进行测序鉴定。方法:利用PCR技术从人全血基因组DNA中扩增出AD启动子片段,经纯化酶切后克隆入载体pGL-3 basic中,形成重组质粒,筛选阳性克隆进行测序,并与GenBank比对。结果:从人全血基因组DNA中PCR扩增得到1.1 kb和2.1 kb AD启动子片段,构建2种报告基因载体,将其命名为AD promoter1.1-pGL-3 basic和AD promoter2.1-pGL-3 basic,经测序证实所插入的目的片段与GenBank检索的人AD启动子序列99.6%匹配。结论:成功构建了含人AD启动子片段的报告基因载体。This study was designed to clone human adiponectin （AD） promoter and construct its luciferase reporter vectors. Total DNA was extracted from human leukocytes and used for amplifying two types of human AD promoter using polymerase chain reaction （PCR） technique. The amplified fragments were inserted into pGL-3 basic vector to construct two types of pGL-3-AD promoter plasmid containing 1.1 kb and 2.1 kb of human AD promoter. The sequence of two recombinant plasmids was determined using gene sequence analysis. The results showed that the recombinant plasmid pGL-3 basic containing human AD promoter was correctly constructed. The sequence was verified to share 99.6% homology with the sequence published at GenBank. It was concluded that the luciferase reporter vector containing human AD promoter is successfully established, which is useful in bioluminescence imaging technology in vitro.

关 键 词：脂联素基因 启动子 克隆人 基因组DNA 荧光素酶报告基因 

分 类 号：R737.250.2[医药卫生—肿瘤]