食品工业科技2011，Vol.32Issue(7)：217-219,236,4.
抗克伦特罗单链抗体在毕赤酵母GS115中的表达
Expression of a single chain Fv antibody against clenbuterol in Pichia pastoris
摘要
Abstract
A Pichia pastoris strain expressing recombinant anti-clenbutero (CBL) single chain Fv (scFv) was constructed.The recombinant plasmid,pPICZαA-CBL,was linearized by endonuclease BstX I and transformed into Pichia pastoris GS115 by electroporation.The multicopy transformants were obtained by Zeocin hyper-resistance screening and were induced to express by methanol.The induced culture supernatant was collected and detected by SDS- PAGE, western bloting and indirect competitive ELISA.The results showed that CBL scFv gene was inserted into the chromosome of GS115 successfully. The CBL scFv expressed from the pPICZαA - CBL recombinants showed 41kDa and anti-His-tag mAb recognized-activity by SDS-PAGE and Western blotting respectively,and could competitively combine with CBL, the IC50, was 5.1ng/mL.The yield of the CBL scFv was 0. 095g/L.lt suggested that a Pichia pastoris strain expressing CBL scFv had been constructed successfully.Further large scale production of CBL scFv could be continued on the basis of our study.关键词
克伦特罗/单链抗体/毕赤酵母/蛋白表达Key words
clenbutero/single chain Fv antibody/Pichia pastoris/protein expression分类
轻工纺织引用本文复制引用
陈舒迟,何永盛,徐小艳,陆田珍,王弘..抗克伦特罗单链抗体在毕赤酵母GS115中的表达[J].食品工业科技,2011,32(7):217-219,236,4.基金项目
国家自然科学基金(30400354) （30400354）
广东省自然科学基金(04300502). （04300502）
