适用于甜玉米叶片蛋白质组分析的双向电泳技术

采用改良的TEA／丙酮法提取叶片总蛋白，建立了一套适用于甜玉米叶片的蛋白质组学研究方法，并对蛋白溶解液、蛋白上样量、第一向IEF等电聚焦时间等条件进行了优化．用改进的蛋白溶解液提取叶片总蛋白，明显减轻了离子干扰造成的蛋白点横拖；500μg蛋白上样量的检出蛋白点最丰富，可达750个以上；延长8000V等电聚焦至30000Vh可获得良好的蛋白点分离效果；采用优化条件后的双向电泳参数，结果重复性较高，检出蛋白点的线性回归相关系数可达0．93以上．...

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Bibliographic Details
Published in华南农业大学学报 Vol. 33; no. 3; pp. 421 - 423
Main Author 王猛 李云锋 王振中
Format Journal Article
LanguageChinese
Published 华南农业大学资源环境学院,广东广州,510642 2012
Subjects
双向电泳
甜玉米
蛋白质组
甜玉米
双向电泳
蛋白质组
Online AccessGet full text
ISSN1001-411X
DOI10.3969/j.issn.1001-411X.2012.03.030

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Summary:采用改良的TEA／丙酮法提取叶片总蛋白，建立了一套适用于甜玉米叶片的蛋白质组学研究方法，并对蛋白溶解液、蛋白上样量、第一向IEF等电聚焦时间等条件进行了优化．用改进的蛋白溶解液提取叶片总蛋白，明显减轻了离子干扰造成的蛋白点横拖；500μg蛋白上样量的检出蛋白点最丰富，可达750个以上；延长8000V等电聚焦至30000Vh可获得良好的蛋白点分离效果；采用优化条件后的双向电泳参数，结果重复性较高，检出蛋白点的线性回归相关系数可达0．93以上．
Bibliography:sweet corn ; proteomics ; two-dimensional electrophoresis
44-1110/S
A two-dimensional electrophoresis protocol suitable for the separation of proteins from sweet corn leaves was established by using improved TCA/aeetone method for total proteins extraction. Two-dimensional electrophoresis conditions such as lysis buffer, protein loading quantities, isoelectric focusing time were modified and improved. The use of modified lysis buffer can lighten the protein horizontal drag ; the maximum number of spots were larger than 750, acquired at 500 μg protein loading quantities; the best total isoelectric focusing time was 50 000 Vh ; the linear regression correlation coefficient of spots was greater than 0. 93, which showed high experiment repeatability.
WANG Meng, LI Yun-feng, WANG Zhen-zhong （College of Natural Resources and Environment, South China Agricuhural University, Guangzhou 510642, China）
ISSN:1001-411X
DOI:10.3969/j.issn.1001-411X.2012.03.030