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目的对克隆的人肥胖 (OB)基因蛋白表达产物－瘦素 (Leptin)进行分子量及特异性鉴定。方法以商品 Leptin为阳性对照，同时设有未经热诱导的对照组 (OB-c)及克隆人肥胖基因重组表达产物试验组 (OB-h)。本文采用 SDS-PAGE对大肠杆菌所表达的蛋白产物进行分子量测定，同时用 Western blotting试验对其产物进行了特异性鉴定。结果经 SDS-PAGE检测，在 21.5～ 14.4 ku之间有一明显蛋白表达条带，对照组在相同的位置上未见明显蛋白表达。试验组 OB-h蛋白表达产物分子量的位置与商品 Leptin的位置基本一致。经 Western blotting试验，在硝酸纤维素膜上，商品 Leptin阳性对照及 OB-h试验组大约 16 ku的位置上可见到两条明显的与抗人 Leptin多克隆抗体呈特异性结合反应染色条带。结论人的 OB基因克隆后，在大肠杆菌中能特异的表达人的 Leptin蛋白质。
Objective To identify the molecular weight and specificity of cloned human leptin (OB) gene product. Methods Commercially available Leptin was used as a positive control, with no heat-induced control (OB-c) and recombinant human cloned OB-h (OB-h). In this paper, the molecular weight of the protein product expressed by E. coli was determined by SDS-PAGE, and its product was identified by Western blotting. Results SDS-PAGE showed that there was a significant protein expression band between 21.5 and 14.4 ku, while the control group showed no obvious protein expression at the same position. The position of the OB-h protein expression product molecular weight in the test group is basically the same as the position of the commodity Leptin. Western blotting showed that two obvious bands of binding reaction with anti-human Leptin polyclonal antibodies were observed on the nitrocellulose membrane, the commercial Leptin positive control group and the OB-h test group about 16 ku. Conclusion Human OB gene can express human Leptin protein specifically in E. coli.