This work was supported by grants from The National Science Fund of China for Distinguished Young Scholars (30825030), The National Natural Science Foundation of China (90717110), Hi-Tech Research and Development Program of China (2007AA10Z122) and Doctor Fund of Xinjiang Construction Corps (2008JC06)
Auxin response factors (ARFs) are important transcription factors involved in auxin signal transduction pathway. In order to elucidate the function of tomato ARF2, we isolated the SlARF2 gene and analyzed its molecular features, in addition, we observed the subcellular localization of ARF2 in transgenic tomato plants. Physicochemical properties and molecular features of ARF2 were predicted by bioinformatic approaches including physical and chemical properties analysis, hydrophobicity analysis, domain analysis, phylogenetic tree analysis and subcellular localization analysis. Moreover, the full-length of SlARF2 gene was amplified by RT-PCR, and a binary vector consisting of ARF2 fused with the yellow fluorescent protein (YFP) coding sequence was further constructed. Using the method of Agrobacterium-mediated transformation, the recombinant vector was transformed into wild-type tomato, and the transgenic tomato plants were confirmed by PCR. The roots of transgenic tomato seedlings were observed under a fluorescence microscope. Bioinformatic analysis showed that the SlARF2 is an unstable soluble protein, which is rich in Ser, Gly, Pro and Leu residues, and it has typical structure and functional domain of ARF family. Tomato ARF2 shares 70.08 %, 66.94 %, and 60.87 % similarity in amino acid sequences with grapes, cassava and Arabidopsis, respectively. Analyses of restriction enzyme digestion and sequencing confirmed the expression vector pBA-ARF2-YFP was successfully constructed. PCR results showed the fusion protein was expressed in transgenic tomato plants. The subcellular localization of SlARF2 was found mostly in the nucleus. SlARF2 has the key characteristics of transcription factor, which is localized in the nucleus, and it might play a pivotal role in tomato fruit development and ripening. This might provide references for studying the function of SlARF2 gene.
FENG Yuan-Yuan, HOU Pei, LI Ying-Nan, LIU Yong-Sheng. Bioinformatic Analysis and Subcellular Localization of Solanum lycopersicum ARF2[J]. Progress in Biochemistry and Biophysics,2012,39(1):51-58
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