The Expression And Potential Interaction Proteins' Verification Of Rice Cytoplasmic Sterile-related Protein ORF216

Plant cytoplasmic male sterility(CMS) is the plant with male sterility inherited maternally cannot produce functional pollens but normal female gametes. In the early study our lab found CMS-related protein ORF216 might interact with some potential proteins. In this study the prokaryotic expression vectors for HL-CMS genes orfH79, potential genes OsRPLx and OsATP5 x, were constructed and the proteins were induced. The mitochondrial proteins were also extracted from different rice etiolated seedlings and the expression of ORF216. The relationships between ORF216 and the potential interaction proteins were studied by methods of the pull-down and western blot detection. The results were including:1. Using pull-down experiment to study the relationship between the two expression proteins ORF216 and ORFH79. The western blot analysis showed that there is no direct interaction between ORF79 and ORF216.2. Much higher expression of ORF216 in the mitochondria from etiolated seedlings of YTA and HL-2, than in that of 9311 were detected by Western Blot analysis. But no expression in that of YTB was observed in the same detection.3. The direct interaction between the OsRPLx and ORF216 was demonstrated using pull-down and western blot analysis. But there was no direct interaction between ATP5 x and ORF216.