重组猪生长激素抗体的制备与纯化  

Preparation and Purification of Polyclonal Antibodies Against Recombinant Porcine Growth Hormone(pGH)

作　　者：姜秀英[1] 朱江[2] 

机构地区：[1]徐州医药高等职业学校药学教研室,江苏徐州221116 [2]苏州大学生命科学学院,江苏苏州215006

出　　处：《淮阴师范学院学报（自然科学版）》2006年第4期311-313,328,共4页Journal of Huaiyin Teachers College;Natural Science Edition

基　　金：江苏省教育厅自然科学基金资助项目(Q1114020)

摘　　要：为了获得重组猪生长激素抗体,对重组猪生长激素融合基因的真核表达产物进行免疫印迹(Western blot)检测,将利用DNA重组技术构建的重组质粒pPGH020在大肠杆菌(E.coli)BL21中进行诱导表达.表达产物经Ni+亲和层析柱纯化,获得纯化的重组猪生长激素融合蛋白.然后以此为抗原免疫新西兰大白兔,获得多克隆抗体,抗体再经硫铵沉淀、透析和亲和层析纯化.Western印迹结果表明,该纯化抗体显示了良好的免疫反应,其灵敏度比兔抗rpGH抗血清提高50倍,为猪生长激素融合基因在真核细胞的表达及功能鉴定奠定了基础.To obtain the antibody of recombinant porcine growth hormone （rpGH）, which is used to test the product expressed by the fusion gene of recombinant pGH, the author induces the recombinant plastid, which is built by the DNA recombinant technology, to express in E.coli BL21 strain. The expressed product is purified with Ni^＋ affinity chromatographic column so that we obtain the purified fusion protein of recombinant pGH. This fusion protein is used as antigen to immune the male rabbits to obtain the polyclonal antibody. The antibody is purified by sedimentation with （NH4）2SO4, dialysis and affinity chromatographic column. The result of western blot indicates that this purified antibody has a good immune reaction and its sensitivity is higher as 50 times as that of the recombinant pGH antibody of the rabbit. This makes a basis for the function identity and the expression of fusion gene of pGH in eukaryotic cell.

关 键 词：免疫印迹 重组猪生长激素 亲和层析 免疫 多克隆抗体 

分 类 号：Q511[生物学—生物化学]