中文摘要：

以pMD18-T-hucp2为模板，用设计的两对特异性的引物PCR得到hucp2cDNAN端和C端的两断基因片段，然后将其插入到pGEX5x1表达载体上，重组质粒在PCR、双酶切、测序鉴定后，经IPTG诱导，在大肠杆菌BL21表达大量融合蛋白GST-UCP2-N（C），经GST亲和层析纯化蛋白．以该蛋白为抗原免疫ucp2基因敲除小鼠，western blot鉴定血清中抗体的反应性．实验证明成功构建了构建pGEX-5X-1-UCPN（c）重组质粒，表达出了融合蛋白．并获得了抗血清．为进一步获得灵敏性更高，特异性更强的UCP2的抗体打下了基础．

英文摘要：

According to sequence of hucp2, we designed two pairs of primers and insert BamH Ⅰ and Xho Ⅰ restrictive endonuclease sites into 5＇ termital and 3＇termital respectively. Using hucp2 clonal vector as template we obtain Extro-cellular fragment of gene DNA at 5r termital （82-249） and 3＇termital （706-927）, which are named as ucp2-N and ucp2-C, then inserted into expressive vector and transform into E. coll. The recombined vectors were identified by PCR, restrictive endonucleases and sequencing. BL21 expression strain was propagated in lura broth. A series of methods （low temperature induction, shorten of induction time, etc. ） were tried to induce soluble GST UCP2-N（C） fusion proteins but without successes. Fusion proteins were mainly expressed as including body. It has showed that the most high-level expression was after 4 hours induced by 0.5 mmol/L IPTG at 28℃ for 4 hours. Pellet was resuspended in STE buffer with 0.03% SDS before sonieation. 0. 1% TritonX-100 was added into supernatant. Fusion proteins were purified by GST affinity chromatography western blot showed a single band at 33 kD. After Emulsifying purified fusion protein in complete Freund＇s adjuvant, we immuned ucp2-KO mice through intraporatroneal injection. 4 weeks later, we strengthened immune response by injecting three times every 2 weeks with fusion protein emulsified in incomplete Freund＇s adjuvant. Blood collected from tails was centrifuged, using theserum we got as the first antibody western blot has done to identify the serum. It resulted two bands at and behind 34kD. The result above indicated that GST-UCP-N（C） was expressed productively, but the concentration of fusion proteins were low without including body denaturation, refolding which should do to improve the concentration of fusion protein.