IL-2与绿脓杆菌外毒素A嵌合蛋白的研究  

Preparation of IL-2/Pseudomonas Exotoxin A Chimeric Protein

作　　者：刘柱[1] 周华[1] 姚为民[1] 万里[1] 张雪梅[1] 王志武[2] 石晓丽[1] 郑辉[1] 朱宏伟[1] 盛君[1] 周雪艳[3] 陈宇[3] 贺玉泉[3] 周传开 

机构地区：[1]长春生物制品研究所,长春130062 [2]白求恩医科大学 [3]中日联谊医院,长春130021

出　　处：《中国生物制品学杂志》2000年第1期16-18,共3页Chinese Journal of Biologicals

摘　　要：目的获得ＩＬ－２绿脓杆菌外毒素Ａ嵌合蛋白，用于相关疾病的治疗。方法用ＰＣＲ方法扩增分 离ＩＬ－２和ＰＥＡ的基因，将ＩＬ－２基因的３’端与绿脓杆菌外毒素Ａ基因的５’端连接后克隆至ｐＢＶ２２０上，转化大肠杆菌 ＤＨ５ａ。挑取菌落培养，快速提取质粒进行酶切，并在温控条件下表达。结果经鉴定，获得重组质粒ｐＢＶ／ＩＬ－２／ ＰＥＡ。温控诱导表达后ＳＤＳ－ＰＡＧＥ分析表明，在５１ ０００处有蛋白表达带。以鼠抗人ＩＬ－２单克隆抗体经免疫印迹验 证，该表达蛋白为所设计的嵌合蛋白。结论本研究建立了一种制备ＩＬ－２／ＰＥＡ嵌合蛋白的方法，该方法也可用于 表达其他蛋白。Objective To prepare -2/Pseudomonas exotoxin A (PEA) chimeric protein used for the treatment of related diseases. Methods After IL-2 and PEA genes were amplified by PCR, the 3' terminal of IL- 2 gene and 5' terminal of PEA gene were linked, then cloned into vector pBV220 and transformed to E. coli DH5a. The colonies were selected out and cultured, then plasmid was extracted rapidly , digested with enzyme, and expressed under temperature control. Results A recombinant plasmid pBV/IL-2/PEA was constructed. SDS-PAGE showed that the molecular weight of expressed protein was 51 000. Western blot with anti-human IL-2 McAb showed that the expressed product was designed chimeric protein. Conclusion A method for preparing IL-2/PEA chimeric protein was established. The method can also be applied for the expression of other proteins.

关 键 词：绿脓杆菌外毒素A IL-2 基因重组 克隆 嵌合蛋白 

分 类 号：Q936[生物学—微生物学]