GGPPS多克隆抗体的制备与鉴定

Preparation and Identification of Polyclonal Antibodies of GGPPS

构建人香叶基香叶基二磷酸合成酶基因(GGPPS)原核表达载体,制备兔抗人GGPPS多克隆抗体.利用DNA重组技术构建原核表达载体pGEX-4T-GGPPS,诱导表达GGPPS融合蛋白,分离纯化该融合蛋白后,作为免疫原免疫家兔制备多克隆抗体.成功构建了pGEX-4T-1-GGPPS原核表达载体,转化Rosetta菌株后可高效表达GGPPS融合蛋白,成功制备GGPPS多克隆抗体,间接ELISA法测定抗体效价为1∶32000,并且抗体具有良好特异性.成功克隆GGPPS基因并制备了特异性的兔抗人GGPPS多克隆抗体,为进一步研究GGPPS的功能奠定了基础.

To construct the prokaryotic expression vector of human GGPPS gene and prepare the specific polyclonal antibody against GGPPS. Construct the prokaryotie expression vector of pGEX- 4T- GGPPS by DNA recombination technology and induce the expression of GGPPS fusion protein. The fusion protein was purified and injected into rabbits to produce polyclonal antibody. The prokaryotie expression vector of pGEX-4T-GGPPS was constructed and the antibody of GGPPS was obtained. The titer of the antiserum was 1：32 000. The specific polyclonal antibody against GGPPS was successfully prepared. The polyelonal antibodies preparation against GGPPS provided a good tool for us to study the function involvement of GGPPS.