Cloning and Sequencing of Human Leptin Gene cDNA

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.