鸡Perilipin1抗血清制备及组织表达特性分析  

Preparation of Antiserums against Chicken Perilipin1 and Tissue Expression Analyses of Perilipin1

作　　者：王彦博[1] 王宁[1] 王丽[1] 王宇祥[1] 李玉茂[1] 李辉[1] 

机构地区：[1]东北农业大学动物科技学院,哈尔滨150030

出　　处：《畜牧兽医学报》2011年第3期349-355,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点基础研究发展计划(973计划)项目(2006CB102105);国家重点基础研究发展计划(973计划)项目(2009CB941604);国家高技术研究发展计划(863)项目(2006AA10A120)

摘　　要：通过制备鸡脂滴包被蛋白(Perilipin1)的抗血清,分析鸡Perilipin1的组织表达特性并推测其在脂肪组织脂类代谢过程中发挥的功能。利用RT-PCR的方法扩增鸡Perilipin1基因,并将其构建到含有His标签的表达载体pET-28a上。将阳性重组表达质粒pET-28a/Perilipin1导入大肠杆菌BL21,在IPTG诱导下产生His/Perilipin1融合蛋白,对产生的His/Perilipin1蛋白包涵体进行复性处理,复性后以其为免疫原制备鸡Perilipin1多克隆抗血清。通过ELISA和Western blot的方法检测抗血清的效价及特异性,并利用此抗血清分析鸡Perilipin1在各种组织中的表达特性以及比较其在高、低脂系公鸡间的表达差异。ELISA和Western blot结果表明,本试验获得了效价高、特异性强的鸡Perilipin1抗血清,同时Western blot检测Perilipin1的组织表达特性结果表明,Perilipin1仅在鸡脂肪组织中表达,而在肝脏、十二指肠、胸肌、腿肌、肌胃、心脏、脾脏、肾脏等组织中未检测到表达信号;Perilipin1在肉鸡高脂系脂肪组织中的表达量显著高于低脂系中的表达量(P<0.05)。结果表明,Perilipin1的表达与肉鸡腹部脂肪的沉积具有密切的关系。The aim of this study was to analyze the expression characteristics of chicken Perilipin1 in different tissues and the function of Perilipin1 in lipid metabolism by preparing the antiserum against chicken Perilipin1.Chicken Perilipin1 gene was amplified by RT-PCR and inserted into pET-28a vector.The recombinant His/Perilipin1 was expressed in E.coli with IPTG induction and renatured by Protein Refolding Kit.The renatured His/Perilipin1 was used as immunogen to immunize the rabbit for preparing polyclonal antibody.The titer and specificity of this antibody were detected by ELISA and Western blot,and the expression characteristics of chicken Perilipin1 in different tissues was analyzed by Western blot.ELISA and Western blot results showed that the high titer and specificity of the antiserum against Perilipin1 was obtained.The tissue expression characterization analysis by Western blot showed that Perilipin1 only expressed in chicken adipose tissue,but no signal was detected in liver,duodenum,breast muscle,leg muscle,gizzard,heart,spleen and kidney.Furthermore,the Perilipin1 expression characteristics between fat and lean broiler lines were also detected,and the results showed that Perilipin1 expressed significantly higher in fat line broilers adipose tissue than that in lean ones（P0.05）.These results of this study suggest that there is a close relationship between the expression of Perilipin1 and the deposition of abdomen adipose in fat and lean broiler lines.

关 键 词：鸡 脂滴包被蛋白 抗血清 表达特性 

分 类 号：S831.2[农业科学—畜牧学]