文献类型：期刊文献

中文题名：谷胱甘肽转硫酶多克隆抗体的制备及应用

英文题名：Preparation and Application of Polyclonal Antibodies against GST

作者：徐猛[1];卫军[2];严明[2];李臻[2];张祖传[2];杨曜中[1]

机构：[1]华东理工大学生物工程学院,上海200237;[2]中国科学院上海生物化学与细胞研究所,上海200031

年份：2002

卷号：28

期号：2

起止页码：153

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;EI(收录号：2002397116935);Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：谷胱甘肽转硫酶;多克隆抗体;免疫印迹;免疫亲和层析;融合表达

外文关键词：GST; polyclonal antibodies; western blot; immunoaffinity chromatography; fusion expression

摘要：将含有谷胱甘肽转硫酶 ( GST)基因的 p GEX质粒转入大肠杆菌 TG1后 ,在 0 .5 mmol/L异丙基硫代 - β- D-半乳糖苷 ( IPTG)诱导培养下表达了 GST,用亲和纯化得到的 GST免疫新西兰大白兔 ,获得了滴度 ( ELISA)在 2× 1 0 5以上的抗血清 ,纯化后的抗体已成功地应用于两种新的融合蛋白 GST- FLP和 GST- Pre
Glutathione S transferase(GST) gene fusion system has being widely used in genetic engineering for expression of foreign proteins. Expressed fusion proteins can be easily detected and purified from bacterial lysates by antibodies against GST. pGEX plasmid vectors which contain GST gene are transformed to E.coli (TG1) for expression of GST in LB medium by inducing of 0.5mmol/L IPTG. The expressed GST was used to immunize rabbits after purification. The title of antiserum against GST after the third immunization was found more than 2×10 5. The preparation of antibodies has been successfully used in Western blot analysis and immunoaffinity chromatography of two novel GST fusion proteins(GST FLP,GST PreS1).