重组靶向毒素IL-6(23)-PE40_(KDEL)在毕赤酵母中的表达及其活性  

Expression of Recombinant Target Toxin IL-6(23)-PE40_(KDEL) in Pichia pastoris and Activity of Expressed Product

作　　者：郭德军[1] 柳增善[2] 潘风光[2] 

机构地区：[1]黑龙江八一农垦大学食品学院,大庆163319 [2]吉林大学人兽共患病研究所细菌室,长春130062

出　　处：《中国生物制品学杂志》2008年第6期471-474,共4页Chinese Journal of Biologicals

摘　　要：目的利用毕赤酵母表达系统表达IL-6(23)-PE40KDEL重组靶向毒素,并检测其杀伤肿瘤细胞的活性。方法采用PCR技术,将目的基因IL-6(23)-PE40KDEL插入到pPIC9载体中SnaBⅠ与NotⅠ位点,电转化至巴斯德毕赤酵母GS115中,筛选Mut-型重组酵母;甲醇诱导表达,体外细胞试验检测其细胞毒性。结果获得12株Mut-重组酵母,其中8株具有细胞毒性,表达产物占上清液蛋白的9%～12.7%;15"l以上的表达产物在体外对SP2/0、HL-60、HepG2、BGC-832和HCT-116细胞具有高度杀伤活性,对CK、HeLa和NIH/3T3细胞无明显影响。结论IL-6(23)-PE40KDEL重组靶向毒素在毕赤酵母GS115中获得表达,并具有选择杀伤肿瘤细胞的活性。Objective Abstract Objective To express recombinant target toxin IL-6 （23）-PE40KDEL in Pichia pastoris and determine the killing activity of expressed product to tumor ceils. Methods Amplify IL-6 （ 23 ）-PE40KDEL gene from the previously constructed recombinant plasmid pKK-IL-6 （ 23 ）-PE40KDEL by PCR, and insert to the SnaB Ⅰ and Not Ⅰ sites of vector pPIC9. Transform the constructed recombinant plasmid pPIC0-SN to P. pastoris GS115 by electroporation, screen Mut-recombinants for expression under induction of methanol. Determine the cytotoxicity of expressed product by in vitro cell test. Results A total of 12 Mur recombinants were screened, of which 8 showed cytotoxicity. The expressed product contained 9. 0% - 12. 7% of protein in supernatant, and showed highly killing activity to SP2/0, HL-60, HepG2, BGC-832 and HCT-116 cells, but no significant effect on CK, HeLa and NIH/3T3 cells. Conclusion Recombinant target toxin IL-6（ 23 ）-PE40KDEL was expressed in P. pastoris and showed selectively killing activity to tumor cells.

关 键 词：重组靶向毒素 毕赤酵母 表达 活性 

分 类 号：Q786[生物学—分子生物学]