Subcellular localization of proteins of Oryza sativa L. in the model tobacco and tomato plants |
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| Authors: | Venugopal Rao Kokkirala Yonggang Peng Sadanandam Abbagani Zhen Zhu Pavan Umate |
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| Institution: | 1.Department of Biotechnology; Kakatiya University; Warangal, India;2.State Key Laboratory of Plant Genomics; Institute of Genetics and Developmental Biology; Chinese Academy of Sciences; Beijing, China;3.National Plant Gene Research Center; Institute of Genetics and Developmental Biology; Chinese Academy of Sciences; Beijing, China |
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| Abstract: | The cellular localization and molecular interactions are indicative of functions of a protein. The development of a simple and efficient method for subcellular localization of a protein is indispensable to elucidate gene function in plants. In this study, we assessed the feasibility of Agrobacterium-mediated transformation (agroinfiltration) of tobacco and tomato leaf tissue to follow intracellular targeting of proteins from rice fused to green fluorescent protein (GFP). For this, a simple in planta assay for subcellular localization of rice proteins in the heterologous host systems of tobacco and tomato leaf via transient transformation was developed. We have tested the applicability of this method by expressing GFP fusions of the putative antiphagocytic protein 1 (APP1) (OsAPP, LOC_Os03g56930) and ZOS3-18-C2H2 zinc-finger protein (OsZF1, LOC_Os03g55540) from Oryza sativa L. subsp. japonica in tobacco and tomato leaf tissues. Our results demonstrate the suitability of GFP as a reporter in gene expression studies in tomato cv. MicroTom. The use of GFP-fused proteins from rice for subcellular targeting in the heterologous hosts of tobacco and tomato plant systems has been confirmed.Key words: agroinfiltration, confocal microscopy, GFP fusion protein, tomato cv, microtom |
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