【摘要】 利用IPTG诱导含有原核表达质粒pGEX-4T-1/PPARγ和pGEX-4T-1/C/EBPα的大肠杆菌BL21(DE3)菌株,表达GST-PPARγ和GST-C/EBPα融合蛋白,表达产物用Glutathione Sepharose 4B纯化,将纯化后的重组蛋白免疫肉仔鸡,检测其产生抗体情况及其对肉鸡腹脂含量的影响。SDS-PAGE结果显示融合蛋白GST-PPARγ在大肠杆菌胞内主要以包涵体的形式表达,分子量大约80ku;融合蛋白GST-C/EBPα在大肠杆菌胞内主要以可溶性蛋白形式表达,分子量大约62ku。主动免疫肉鸡结果表明PPARγ、C/EBPα及混合组鸡只均产生了较高水平的抗体,且PPARγ组肉鸡腹脂含量显著低于对照组。更多还原

【Abstract】 In the current study,fusion protein GST-PPARγ and GST-C/EBPα were expressed in E.coli. BL21(DE3) which has prokaryotic expression plasmids pGEX-4T-1/PPARγ and pGEX-4T-1/C/EBPα with IPTG induction. The expressed fusion proteins were purified by Glutathione Sepharose 4B affinity chromatography,and then were used to immunize broilers for analysis the levels of antibody of PPARγ and CEBPα and the effects on abdominal fat contents in broiler. SDS-PAGE showed that the GST-PPARγ fusion protein formed mostly inclusion body in E.coli. and its molecular weight was about 80 ku,and the GST-C/EBPα fusion protein was expressed in E.coli. with mostly soluble-form protein and its molecular weight was about 62 ku. Inactive immunization results showed that there were high levels of antibody against PPARγ and C/EBPα in serum of broiler,and the broilers of PPARγ group had significant lower abdominal fat contents than control group.更多还原