人瘦素基因克隆与序列测定  

Cloning and Sequencing of Human Leptin Gene cDNA

作　　者：徐明彤[1] 吕凌[2] 钟光恕[3] 程桦[1] 傅祖植[1] 

机构地区：[1]中山医科大学孙逸仙纪念医院内分泌科 [2]中山医科大学分子医学研究中心,广东广州510120 [3]孙逸仙纪念医院内分泌科

出　　处：《中山医科大学学报》2000年第2期104-107,共4页Academic Journal of Sun Yat-sen University of Medical Sciences

基　　金：广东省自然科学基金! ( 95 0 3 64 )

摘　　要：【目的】构建人瘦素基因cDNA克隆 ,并进行序列分析。【方法】用逆转录聚合酶链反应扩增人瘦素基因cDNA ,获得片段 (6 40bp)连接至 pUC19载体 ,并转化大肠杆菌DH5α。以末端终止法进行序列测定。【结果】所克隆的人瘦素cDNA含全长的人瘦素基因编码区 ,仅 2 87位的腺苷酸被鸟苷酸代替 ,导致 96位编码谷氨酰胺的密码子CAA改变为编码精氨酸的CGA ,其意义尚待阐明。【结论】以逆转录聚合酶链反应方法成功地构建了人瘦素基因cDNA克隆。Objective To construct the cDNA clone and analyze the sequence of human leptin gene. Methods The full human leptin cDNA fragment was amplified by reverse transcription polymerase chain reaction (RT PCR). The obtained fragment of 640 bp was inserted into pUC19 vector and transformed to E. Coli DH5α. The recombinant plasmid pUC19 ob was sequenced by dideoxy nucleotide chain termination method. Results A full human leptin cDNA fragment was obtained, and a base substitution (A to G) of coding region at the position 287 was detected leading to a replacement of glutamin by arginine. Its physiological significance need further studying. Conclusions The human leptin gene cDNA clone was constructed by RT PCR.

关 键 词：瘦素 DNA序列测定 基因克隆 

分 类 号：Q785[生物学—分子生物学]