β_2肾上腺素受体基因在毕赤酵母中的表达及活性测定  

Expression of Functional Mouse β_2-adrenergic Receptor in the Methylotrophic Yeast Pichia pastoris and Activity Determination

作　　者：黄园园[1] 沈红[1] 王迪[2] 于洪侠[2] 孙丰梅[2] 杨啸枫[2] 杨曙明[2] 

机构地区：[1]北京农学院,北京102206 [2]中国农业科学院饲料中心,北京100081

出　　处：《中国生物工程杂志》2008年第8期18-22,共5页China Biotechnology

基　　金：科技部"科研院所社会公益科研专项"(2005DIB4J040)资助项目

摘　　要：为提高β2-肾上腺素受体(β2AR)表达量,满足生产需求,使其应用到抗体技术上,利用分子克隆技术将β2-肾上腺素受体基因(β2AR)与绿色荧光蛋白基因(eGFP)克隆到毕赤酵母(Pichia pastoris)表达载体中。表达载体pPICZαDNAsGFP转化至酵母后,β2AR和eGFP基因与酵母基因重组。利用筛选出的阳性重组子诱导表达β2AR和eGFP的融合蛋白,通过125I标记的配体结合实验证明获得了有受体活性的融合蛋白。Object： to obtain large quantities of β2-adrenergic receptor （β2AR） and detect β2-agonist. Method：Mouse β2AR cDNA gene was amplified by PCR, then linked with EGFP and inserted into yeast expression vector pPICZαA. The reconstructed plasmid, PICZαDNAsGFP was transformed into Pichia pastoris KM71. After induction, a recombinant protein about 75kDa in size was determined by Western blot. ^125ICYP was used to determine the activity of recombinant β2AR. These results showed that the expressed protein is active.

关 键 词：Β2肾上腺素受体 EGFP 毕赤酵母表达系统 pPICZαA Β2激动剂 

分 类 号：Q786[生物学—分子生物学]