摘要
目的:实现重组牛促卵泡激素在毕赤酵母中的表达。方法:依据毕赤酵母的密码子偏爱性设计并利用PCR方法合成了牛促卵泡激素的α亚基和带有6×his-tag的β亚基相应的DNA序列,构建表达载体pHIL-S-bFSH,转化毕赤酵母,表达蛋白进行ELISA定量和Western blot鉴定。结果:重组牛促卵泡激素的表达量为0.26mg/L,Western blot鉴定分子量为35kDa。结论:在毕赤酵母中实现了具有免疫源活性的重组牛促卵泡激素的表达。
Objective： To express recombinant bovine follicle stimulating hormone （bFSH） using Pichia pastoris. Methods： Two DNA sequences coding for subunit α and subtmit β with the C- terminal end of 6×his- tag were optimized based on synonymous codons usage preferred by the methylotropic yeast P. pichia. These genes were synthesized based on a polymerase chain reaction （PCR）. The vector pHIL- S - bFSH was constructed and transformed Pichia pastoris. The expressed protein was analyzed using ELISA and Western blot. Results： The expression level of recombinant bFSH was 0.26mg/L, the molecular was 35kDa. Conclusion： The recombinant bovine FSH was successfully expressed using P. pastoris.
出处
《生物技术》
CAS
CSCD
2007年第4期29-31,共3页
Biotechnology
关键词
牛促卵泡激素
毕赤酵母
表达
bovine follicle stimulating hormone
Pichia pastoris
expression
