中国人肥胖基因真核表达载体的构建

为研究肥胖（ｏｂｅｓｉｔｙ）的病因及肥胖基因（ｏｂ）的表达与调控，根据文献报道的ｈｏｂ序列设计引物，经ＲＴ－ＰＣＲ扩增中国人的ｏｂ基因（包括信号肽在内的ｃＤＮＡ全长５４０ｂｐ），ＰＣＲ产物采用Ｔ－Ａ克隆法首先连接到克隆载体ｐＵＣ１１９，然后定向转移到经改造的真核表达载体ｐＳＶ－β－ｌａｃＺ，酶谱分析表达克隆基因为的人肥胖基因。

CONSTRUCTION OF EUCARYOTIC EXPRESSION VECTOR OF CHINESE OBESE GENE

In order to better understand the pathoseny of the obesity and the mechanism of expres sion and manipulation of the obese (ob) gene- we designed and synthesized a pair of specific primers based on the published human ob gene to amplify Chinese ob gene by RT - PCR'The amplified prod uct, \"hich was 54obp in length including the whole DN A sequencescoding for the signal pep1ide and mature peptidc, was firstly 1igated to cloning vectorpU C119 and then directiona1ly transfered to modified eucaryotic expression vector pSV -6lacZ. Enzymatic analysis showed that the cloned gene was human obese gene