Proteomic analysis of transgenic Arabidopsis plants over expressing GR-RBP2 in comparison with Wild Type

Glycine-Rich RNA Binding Protein2 (GR-RBP2) is one of the eight GR-RBP family members in Arabidopsis thaliana which is located in mitochondria after translation. Here we employed a proteomic approach to compare transgenic Arabidopsis plants over expressing GR-RBP2 with Wild Type. In this study، GR-RBP2 gene was isolated from Arabidopsis and cloned into pBIN19 vector. Recombinant vector was transferred to Arabidopsis (Col-0) via Agrobacterium tumefaciens by Floral Dipping method. Transformation with full length cDNA of GR-RBP2 yielded 21 lines. Over expressing plants are indistinguishable from WT seedlings. The whole lines are unchanged with regard to flowering time and are fully fertile. Gene expression levels were determined in different over expressing lines compare with WT seedlings. The mRNA level in over expressing lines was 2 times higher than WT. Proteomic analysis of over expressing lines were carried out on two different gel systems: 2D IEF/ SDA PAGE and 2D Blue-native/SDS PAGE. All gels were silver stained. 2D gel electrophoresis revealed that GR-RBP2 was induced in over expressing lines. Results showed that respiratory chain looks very much in the same investigated lines and WT. Complex V was partially degraded (F1 complex)، but this also is the same in all samples. Approximately 110 spots were identified by mass spectrometry. Six protein spots were increased in over expressing lines compared to WT. A look at some well known proteins showed that there is no major change indicating induction or repression of metabolic pathways.