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Protein extraction and database construction in tea rhizosphere soil
Acta Physiologiae Plantarum ( IF 2.6 ) Pub Date : 2020-09-30 , DOI: 10.1007/s11738-020-03146-5
Hai-bin Wang , Chun-lian Zhu , Yu-hua Wang , Qing-xu Zhang , Peng Wang , Ding Li , Xiao-li Jia , Jiang-hua Ye , Hai-bin He

Soil protein extraction and database construction are the key points of soil proteomics research. In this paper, tea tree rhizosphere soil was used as material. The soil proteins were extracted three times by citrate, SDS, and mixture of citrate and SDS, respectively. The total proteins were separated by 2-DE electrophoresis and identified by Data Dependent Acquisition (DDA). The DDA data collection was further separated by High-Performance Liquid Chromatography (HPLC) and identified by LC–MS/MS, then to build the database of soil protein and microbial species using fungus and bacteria databases. The result showed soil protein was identified and reached 2741 points, and the molecular weight was mainly distributed in between 2.64 and 338.33 kDa, and Isoelectric point (pI) is mainly distributed in between 3.78 and 12.15. The soil protein was mainly from 138 families, 346 species of microorganisms. This optimization method could obtain more proteins than previous methods, with a wider range of molecular weight and pI. This study lays an important foundation for the research and development of soil metaproteomics.



中文翻译:

茶根际土壤蛋白质提取与数据库构建

土壤蛋白质的提取和数据库建设是土壤蛋白质组学研究的重点。本文以茶树根际土壤为材料。用柠檬酸盐,SDS以及柠檬酸盐和SDS的混合物分别提取土壤蛋白3次。通过2-DE电泳分离总蛋白,并通过数据依赖性采集(DDA)鉴定。DDA数据收集通过高效液相色谱(HPLC)进一步分离,并通过LC–MS / MS进行鉴定,然后使用真菌和细菌数据库建立土壤蛋白质和微生物物种的数据库。结果表明,已鉴定出土壤蛋白并达到2741点,分子量主要分布在2.64至338.33 kDa之间,等电点(pI)主要分布在3.78至12.15之间。土壤蛋白主要来自138科346种微生物。与以前的方法相比,这种优化方法可以获得更多的蛋白质,分子量和pI范围更广。该研究为土壤元蛋白质组学的研究和发展奠定了重要的基础。

更新日期:2020-09-30
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