Molecular cloning, sequencing and expression of obese gene in the Chinese

Objective To construct the human obese(ob) cDNA clone in the Chinese, and analyze the expression of the ob gene in adipose tissue of obese, non-obese subjects and nooinsulin-dependent diabetes mellitus (NIDDM) Chinese patients. Methods A ob cDNA clone was isolated by reverse transcription polymerase chain reaction (RT-PCR). Four groups of Chinese subjects participated in the study: 1) 12 obese subjects [body mass index (BMI): 28.5 +/- 2.3 kg/m(2)]; 2) 11 non-obese subjects (BMI: 21.0 +/- 1.5 kg/m(2)); 3) 8 obese NIDDM patients (BMI: 27.0 +/- 1.4 kg/m(2)); 4) 11 non-obese NIDDM patients (BMI: 21.2 +/- 1.4 kg/m(2)). The expression of ob gene mRNA in abdominal subcutaneous adipose tissue was examined using RNA dot blot hybridization with a digoxigenin-labeled human ob cDNA probe. The hybridized signals were quantitated by densitometry. Results A full human ob cDNA fragment which included a glutamine codon at + 49 was obtained. A base substitution (A to G) in the coding region at position 287 was found, resulting in a glutamine being replaced by an arginine. Expression of the ob gene was significantly higher in Chinese obese subjects compared to non-obese ones (P < 0.05) and positively correlated with the BMI. No significant difference in the amount of ob mRNA was detected between non-diabetic and diabetic groups at the same BMI level. Conclusions We constructed a full length human ob cDNA clone. The expression of the ob gene was significantly higher in Chinese obese subjects than in non-obese ones. The metabolic and hormonal changes associated with NIDDM are not the main factors regulating the expression of the ob gene.