大仓鼠β_2肾上腺素受体基因的套叠PCR法多点突变研究  

Multi-mutation of β_2 Adrenergic Receptor Gene from Greater Long-tailed Hamster by Overlap Extension PCR

作　　者：王迪[1] 黄园园[2] 杨曙明[1] 于洪峡[1] 杨啸枫[1] 

机构地区：[1]中国农业科学院农业质量标准与检测技术研究所,中国北京100081 [2]北京农学院,中国北京102206

出　　处：《生命科学研究》2008年第1期45-49,共5页Life Science Research

基　　金：科技部"科研院所社会公益研究专项"项目(2005DIB4J040)

摘　　要：对大仓鼠β2肾上腺素受体蛋白(β2 adrenergic receptor,β2AR)进行立体结构与疏水性分析.利用套叠PCR法对β2AR基因进行多点突变,将保守跨膜域疏水氨基酸突变为正电荷亲水氨基酸,成功构建分别带有7个与3个突变氨基酸的原核突变表达载体.探讨了套叠PCR的反应条件,为β2AR基因在体外的高量表达提供基础.The three-dimensional structure and hydrophobicity of β2AR（β2 adrenergic receptor） were studied. Multi-mutated of greater long-tailed hamster β2 adrenergic receptor gene by the gene splicing by overlap extension PCR（SOE PCR）method which use the positive electrical charge amine acid instead of hydrophobic amine acid on the putative transmembrane helices. Two prokaryotic mutation expression vectors with 7 and 3 mutated amine acids separately were constructed. Over-lap extension PCR reaction condition was discussed. The research was benefit for the high level expressed β2AR protein in vitro.

关 键 词：β2肾上腺素受体基因 G蛋白偶联受体 套叠PCR 突变 

分 类 号：Q782[生物学—分子生物学]