摘要
本研究以籽粒苋根为试验材料,比较3种不同蛋白提取方法对蛋白质双向电泳的影响,并对其中的蛋白上样量和等电点聚焦时间进行了优化。结果表明:与酚抽提法和TCA-丙酮法相比,Tris-Hcl法提取籽粒苋根系蛋白效果较好;上样量800μg和聚焦时间70 000 Vh的条件下会得到理想的双向电泳图谱。采用优化后的条件,可以得到蛋白点数量多、背景清晰的双向电泳图谱,将为下一步镉胁迫下籽粒苋根系蛋白质组学研究及利用分子育种技术培育耐镉的籽粒苋新品种提供技术资料。
In this study,Amaranthus hypochondriacus root was taken as experimental material,root protein sample of A. hybridus prepared by three methods. The protein extraction,suitable concentration of protein per IPG gel and SDS-PAGE parameters of two-dimensional electrophoresis were optimized. The results showed that the protein yield extracted by tris-Hcl method was much higher than that by phenolic extraction and TCA- acetone method. When the sample was 800 μg and focusing time of 70 000 Vh,an ideal two-dimensional electrophoresis map was obtained. The optimized 2-DE system gives the maximum number of protein spots with clear background,which help to the study of proteomics and molecular breeding in A. hybridus under cadmium stress.
出处
《分子植物育种》
CAS
CSCD
北大核心
2015年第8期1884-1889,共6页
Molecular Plant Breeding
基金
国家自科学基金(41101298)资助
关键词
籽粒苋
蛋白提取
双向电泳
Amaranthus hypochondriacus,Protein extraction,Two-dimensional electrophoresis
