Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Education, Northeast Forestry University, 150040, Harbin, China.
Mol Biol Rep. 2010 Feb;37(2):637-41. doi: 10.1007/s11033-009-9478-6. Epub 2009 Mar 14.
Proteins could be well separated and further identified by the use of 2-DE and related techniques. Yet, there are many proteins could not be detected even by more effective dyes because of their inherent low abundance or their low resolution. As a result, polyA-affinity column was used as a method to enrich polyA-binding proteins and then identified by MALDI-TOF-MS. In this study, 23 Arabidopsis chloroplast protein spots coded by 18 genes were identified, and majority of these proteins were classified into three related categories according to their annotations in the Swiss-Prot database, including NAD-, RNA-, and ATP-binding motifs, respectively. The major goal of the present Arabidopsis chloroplast proteomics project was to identify novel polyA-binding proteins or protein isoforms located in Arabidopsis chloroplasts and the specific research of cellular proteins with extremely low transcription levels could be fulfilled.
蛋白质可以通过 2-DE 及相关技术很好地分离和进一步鉴定。然而，由于其固有的低丰度或低分辨率，即使使用更有效的染料，仍有许多蛋白质无法被检测到。因此，聚 A 亲和柱被用作富集聚 A 结合蛋白的方法，然后通过 MALDI-TOF-MS 进行鉴定。在这项研究中，鉴定了 18 个基因编码的 23 个拟南芥叶绿体蛋白斑点，根据它们在 Swiss-Prot 数据库中的注释，这些蛋白质大部分被分为三个相关类别，分别是 NAD-、RNA- 和 ATP-结合基序。本拟南芥叶绿体蛋白质组学项目的主要目标是鉴定位于拟南芥叶绿体中的新型聚 A 结合蛋白或蛋白同工型，并且可以实现对细胞中极低转录水平的蛋白质的特定研究。
