生物技术通报
生物技術通報
생물기술통보
BIOTECHNOLOGY BULLETIN
2010年
4期
103-107
,共5页
非洲山毛豆%叶片%蛋白质组%双向电泳
非洲山毛豆%葉片%蛋白質組%雙嚮電泳
비주산모두%협편%단백질조%쌍향전영
Tephrosia vogelii%Hook f.%Leaf%Proteome%Two-dimensional electrophoresis
以非洲山毛豆叶片为材料,对非洲山毛豆总蛋白质3种提取方法(TCA/丙酮沉淀法、尿素/硫脲法和酚-甲醇/醋酸铵沉淀法)以及3种蛋白裂解液进行比较分析.结果表明,采用酚-甲醇/醋酸铵沉淀法提取非洲山毛豆叶片总蛋白,用蛋白裂解液(7 mol/L尿素,2 mol/L硫脲,4% CHAPS,40 mmol/L Tris-base,1% Bio-Lyte pH3.5-10,65 mmol/L DTT)裂解蛋白1 h,2-DE图谱分离到的蛋白点效果最好.此方法适合于色素、多酚及黄酮类次生代谢物含量较多的非洲山毛豆叶片总蛋白制备方法.
以非洲山毛豆葉片為材料,對非洲山毛豆總蛋白質3種提取方法(TCA/丙酮沉澱法、尿素/硫脲法和酚-甲醇/醋痠銨沉澱法)以及3種蛋白裂解液進行比較分析.結果錶明,採用酚-甲醇/醋痠銨沉澱法提取非洲山毛豆葉片總蛋白,用蛋白裂解液(7 mol/L尿素,2 mol/L硫脲,4% CHAPS,40 mmol/L Tris-base,1% Bio-Lyte pH3.5-10,65 mmol/L DTT)裂解蛋白1 h,2-DE圖譜分離到的蛋白點效果最好.此方法適閤于色素、多酚及黃酮類次生代謝物含量較多的非洲山毛豆葉片總蛋白製備方法.
이비주산모두협편위재료,대비주산모두총단백질3충제취방법(TCA/병동침정법、뇨소/류뇨법화분-갑순/작산안침정법)이급3충단백렬해액진행비교분석.결과표명,채용분-갑순/작산안침정법제취비주산모두협편총단백,용단백렬해액(7 mol/L뇨소,2 mol/L류뇨,4% CHAPS,40 mmol/L Tris-base,1% Bio-Lyte pH3.5-10,65 mmol/L DTT)렬해단백1 h,2-DE도보분리도적단백점효과최호.차방법괄합우색소、다분급황동류차생대사물함량교다적비주산모두협편총단백제비방법.
In this study,three protocols for total protein extraction in Tephrosia vogelii Hook f.leaf,trichloroacetic acid/acetone precipitation (TCA),phenol extraction methanol/ ammonium acetate precipitation and urea/thiourea extraction and three protein lysis buffers were compared.The results showed that,total proteins of samples were extracted using phenol extraction methanol/ ammonium acetate precipitation method and dissolved with the lysis buffer A(7 mol/L urea,2 mol/L thiourea,4% CHAPS,40 mmol/L Tris-base,1% Bio-Lyte pH3.5-10,65 mmol/L DTT)for one hour.The proteins were well separated and the satisfaltory electrophoresis map was obtained.This protein extraction method is suitable for Tephrosia vogelii leaf containing pigments,polyphenol and flavonoids secondary metabolites.
