玉米CMS-S恢复基因Rf_3近等基因系的蛋白质分析

Preliminary Protein Analysis of Near Isogenic Line (NILs) for Rf3 in CMS-S Maize

【目的】Rf3是玉米CMS-S型不育系两个以上恢复基因中的标准基因,对其进行分子生物学研究意义很大。【方法】利用自主创建的Rf3近等基因系不育群体(Ps)和可育恢复系群体(Pf)为试材,采用改进的双向(二维)聚丙烯酰胺凝胶电泳(2D-PAGE)技术,对苗期、成株期叶片细胞总蛋白进行分析。【结果】分离到在Pf遗传背景下分子量为25.2kD,等电点为5.0的特异蛋白质RRPⅡ,优化了2D-PAGE技术。【结论】该特异蛋白质点的出现与消失可能与玉米CMS-S的育性恢复有一定关系,对其纯化分离将为Rf3基因的克隆奠定基础和进一步了解玉米质核互作不育表达模式的分子机制。结合本研究,文章还讨论了采用2D-PAGE技术在玉米分子生物学研究中的优化以及基因组学与蛋白组学研究的衔接。

[Objective] The Rf3 gene is one of two or more nuclear genes required for fertility restoration at the male-sterile S-cytoplasm （CMS-S） male in maize. It is one of the most important research fields in molecular biology. [Method] The near isogenic lines, Ps and Pf, were developed by anther, which serves as materials for isolating and cloning Rf3 gene by using of two-dimensional polyacrylamide gel electrophoresis （2D-PAGE） we had improved in this study. [Result] The results showed that there was a special protein RRP II （MW25.2 kD, pI5.0） in the leaves of Pf during the period of the meiosis of pollen mother cell. [Conclusion] The existence and disappearance of this protein spot may have same relation to the fertility restoration in CMS-S maize. The further isolation and purification of this protein will be beneficial to clone Rf3 gene and discover the molecular mechanism of fertility expressional model. Using the 2D-PAGE system we optimized and trends and directions of the joint studies for genomics and proteomics were discussed here.