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.
出处
《生殖与避孕》
CAS
CSCD
北大核心
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)
关键词
截短型
猪卵透明带-3β
人肠杆菌
表达
发酵
trtmcated
procine zone pellucida
E.coli
gene expression
fermentation
