南京林业大学学报（自然科学版）2017，Vol.41Issue(3)：51-56,6.DOI:10.3969/j.issn.1000-2006.2017.201604047
适于双向电泳分析的银杏叶绿体蛋白质提取方法
An efficient extraction method of chloroplast proteins from ginkgo plants compatible for two-dimensional electrophoresis
摘要
Abstract
[Objective] To explore the mechanism of the response of photosynthesis in ginkgo plants at protein level to environmental conditions,an extraction method for chloroplast proteins was investigated.[Method] An experimental system was developed that was compatible with two-dimensional electrophoresis (2-DE) of ginkgo chloroplast proteins.Trisphenol method was used for the extraction of ginkgo chloroplast proteins,and compared with trichloroacetic (TCA)-acetone precipitation method.[Result] The results indicated that Percoll step gradient centrifugation was suitable for ginkgo chloroplast isolation,and the mean intact rate of purified chloroplasts was approximately 85％.The results of one-dimensional PAGE indicated that protein bands of chloroplasts were more distinct at the low molecular weight region of the gel for the Tris-phenol extracti method compared with the TCA-acetone precipitation method.Moreover,comparison of this method with TCA-acetone precipitation for the extraction of ginkgo chloroplast proteins showed that the method resulted in higher protein yields and produced more protein spots on 2-DE gels with less streak interference.[Conclusion] The above results revealed that the Tris-phenol extraction method could efficiently extract high-quality chloroplast proteins for 2-DE,and could be applied in proteomic analysis of ginkgo chloroplasts.关键词
银杏/叶绿体蛋白质组/蛋白质提取方法/双向电泳Key words
ginkgo/chloroplast proteomics/protein extraction method/two-dimensional electrophoresis分类
农业科技引用本文复制引用
施大伟,何梅,梁霏霏,徐鸣,谢寅峰..适于双向电泳分析的银杏叶绿体蛋白质提取方法[J].南京林业大学学报（自然科学版）,2017,41(3):51-56,6.基金项目
国家自然科学基金项目(31300572) （31300572）
江苏省大学生实践创新训练计划(201410298089X) （201410298089X）
江苏高校优势学科建设工程资助项目(PAPD) （PAPD）
