中文摘要：

瘦素（leptin）由ob基因编码，对调节能量代谢起着重要作用。本研究建立了高原鼠兔瘦素的原核表达系统，并对其进行了原核表达。研究中作者从高原鼠兔瘦素基因的cDNA文库中，扩增编码高原鼠兔瘦素的核酸序列，并利用DNA基因重组技术将其克隆到原核表达载体pE330a（＋）中，构建了高原鼠兔瘦素原核表达载体pET30a（＋）／ppleptin。对目的片段进行测序确认后，将其转化到大肠杆菌B121中，并利用IPTG诱导外源性目的蛋白表达。表达的包涵体蛋白经溶解及变性后上柱纯化。重组质粒经测序检测后，表明原核载体构建正确。同时，SDS—PAGE凝胶电泳结果显示，重组菌在16KD处有明显新增条带，纯化后的目的蛋白条带纯度较高。该结果为高原鼠兔瘦素的后续基础研究提供了基础资料。

英文摘要：

Leptin, the production of the ob gene, plays an important role in the regulation of energy homeostasis. Plateau pika（ Ochotona curzoniae）leptin is closed to cold tolerance. In this study, we establish an effective method for expression of recombinant pika leptin in Escherichia coli. The gene sequence encoding pika leptin was obtained by PCR from the plateau pika cDNA library and the PCR product was cloned into pET30a（ ＋ ） by DNA recombination techniques. After DNA sequencing, the confirmed recombinant clone pET30a（ ＋ ）/ppleptin was transformed into BL21 （DE3） for expression under the induction of IPTG. Due to the expressed protein,the insoluble inclusion body must be separated, denaturated and purified with a Ni sepharose column. The sequencing results of pET30a（ ＋ ）/ppleptin vector demonstrated that the insert of the pika leptin gene was the same as that of pika gene in GeneBank. At the same time the recombinant protein was identified by SDS -PAGE, and the results revealed that there was a new band of protein around 16KD;this protein was purified success- fully. Our results showed that the prokaryotic expression system of pika leptin has been successfully constructed and the purified recombinant protein provides a basis for further research of plateau pika leptin.