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.

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。