基因重组大肠杆菌表达截短型pZP3β的发酵条件优化  

Studies on Expression and Fermentation of Truncated Porcine Zona Pellucida 3β in E.coli

作　　者：谢秋玲[1] 陈小佳[2] 朱伟杰[3] 赵振云[4] 吴建虹[1] 徐万祥[5] 刘侃[1] 李菁[6] 

机构地区：[1]暨南大学生物工程研究所,广州510632 [2]广东省生物工程药物重点实验室,广州510632 [3]暨南大学生殖免疫研究中心,广州510632 [4]暨南大学生物系,广州51063 [5]上海市计划生育科学研究所,上海200032 [6]暨南大学医学院,广州510632

出　　处：《生殖与避孕》2006年第4期195-199,213,共6页Reproduction and Contraception

基　　金：广东省科技计划项目(2001C12001)

摘　　要：目的:对已构建好的表达pZP3β蛋白(pZP130)工程菌的培养条件进行优化,通过在5L罐中发酵和Ni柱纯化,以期获取较大量目的产物。方法:在摇瓶中改变不同的培养及诱导条件,比较工程菌的生长和目的蛋白的表达,优化发酵条件;并在5L发酵罐中进行发酵,获得菌体,破碎后进行Ni柱纯化。结果:优化的发酵条件是:10%的接种量,加入2%甘油的培养基培养菌体3h后加入0.5mmol/LIPTG,诱导4h后收样。在5L发酵罐中发酵,获得菌体量36g/L,经Ni柱纯化,获得蛋白粗制品33mg/L发酵液。结论:截短型pZP3β蛋白可以在E.coli中获得较高表达,并且可以通过发酵和Ni柱纯化得到较大量的蛋白,这有助于pZP的深入研究和应用。Objective： To obtain the recombinant truncated procine zone pellucida protein 3β（tunc-pZP3β） for the further research by expressing the target protein in E.coli in the optimum fermentation conditions. Methods： The fermentation conditions for E.coli BL21（DE3）plyS/pET3c-tp ZP3β were optimized in flasks and the engineered E.coli was fermented in 5 L fermenter. Then the target protein was purified with Ni-chromatography. Results： The optimum fermentation conditions were： incoulum concentration was 10%; 0.5 mmol/L IPTG was added to induce the exogenous protein to express for 4 h aider culturing in LB medium with 2% glycerol for 3 h, We got the biomass of 36 g/L and the recombinant tp ZP3β of 33 mg/L in 5 L fermenter. Conclusion： Under the conditions we optimized, the new truncated protein can meet its high expression in E.coli and was abtained with fermentation and purification. This truncated protein could benefit further investigation for its immunogenicity and the development of antigen preparation.

关 键 词：截短型 猪卵透明带-3β 人肠杆菌 表达 发酵 

分 类 号：Q78[生物学—分子生物学]