拟南芥胱硫醚-γ-合成酶(D-AtCGS)抗血清制备及对D-AtCGS转基因大豆的检测  

Preparation of Polyclonal Antibody against D-AtCGS and Detection of D-AtCGS in Transgenic Soybean Plants

作　　者：王义鹏[1,2] 于洋[3] 孙石[3] 许艳丽[1] 侯文胜[3] 

机构地区：[1]中国科学院东北地理与农业生态研究所,黑龙江哈尔滨150040 [2]东北林业大学生命科学院,黑龙江哈尔滨150040 [3]中国农业科学院作物科学研究所国家农作物基因资源与基因改良重大科学工程,北京100081

出　　处：《大豆科学》2011年第4期537-540,共4页Soybean Science

基　　金：国家重大科技专项资助项目(2008ZX08010-004,2008ZX08004-003)

摘　　要：构建了N端缺失拟南芥CGS基因(D-AtCGS)的原核表达载体,对其进行了原核表达、蛋白纯化和抗血清制备,并利用获得的抗血清对国家大豆改良中心北京分心中心获得的组成型表达、种子特异性表达的D-AtCGS转基因大豆及转全长AtCGS(F-AtCGS)和D-ATCGS的豆科模式植物百脉根进行了Western blot检测,结果表明:各供试转基因材料的检测信号明显高于野生型对照,最终成功建立了AtCGS转基因大豆的Western blot检测方法,该研究可为AtCGS的表达分析和转基因植物检测提供良好的技术支持。Methionine（Met） has very important physiological functions as an essential sulfur-containing-amino acid.Cystathionine-γ-synthase（CGS） is the key enzyme in Met synthesis.In this study,prokaryotic expression vector of Arabidopsis D-AtCGS gene was constructed,and D-AtCGS protein was purified to prepare polyclonal antibody against D-AtCGS.Then transgenic soybean constitutive expressing or seeds specific expressing D-AtCGS and Lotus corniculatus transgenic plants with F-AtCGS or D-AtCGS obtained by Beijing Branch Center of National Center for Soybean Improvement were detected by Western blot method using polyclonal antibody we prepared.Results showed that detection signal of transgenic plants were all much stronger than wild type.At last,Western blot method for detection of transgenic soybean plants with AtCGS gene was successfully established.This work provides technology methods for analysis of CGS expression and detection of CGS transgenic plants.

关 键 词：D-AtCGS 原核表达 抗血清制备 转基因大豆检测 

分 类 号：S565.1[农业科学—作物学]