番茄SlCAT2基因的克隆及功能研究-生物学专业论文.docxVIP

重庆大学 重庆大学硕士学位论文 英文摘要 PAGE PAGE VI ABSTRACT Amino acid transporters, which mediate the transport of amino acids, play an important role in plant growth and development. Based on sequence similarities, the plant transporters can be classified into two major superfamilies: the ATFs (the amino acid transporter family) and the APCs (the amino acid polyamine choline transporter). The CATs family (cationic amino acid transporters) is subfamily of the APCs. The APCs are poorly studied in plant and only a few members of the CATs family have been studied. In this paper, we cloned SlCAT2a and SlCAT2b gene from tomoto, studied their gene sequence and the protein characteristics. We analysed the phylogenetic relations of CATs famiy proteins and the subcellular localizations of SlCAT2a. We also analysed their temporal and spatial expression patterns. More, we constructed the overexpression vectors SlCAT2a and SlCAT2b and the antisense vector AsCAT2. The plant expressive vectors were transformed into tomato by Agrobacterium-mediated method. In addition, we studied the substrate specificity of SlCAT2a and SlCAT2b proteins to investigate the potential physiological function of them in plant growth and development. The main results are as follows: Based on gene sequence homology, two possible EST sequences of tomato SlCAT2 protein were found from tomato genome database (SGN), named SlCAT2a and SlCAT2b. We found their 1476bp were the same and only the 5 nucleotide sequence exsited difference. In addition, SlCAT2a and SlCAT2b were located on chromosome 10, the position of 10 exons on the genome were the same. We cloned SlCAT2a and SlCAT2b using RT-PCR. The result showed that the cloned full-length cDNA of SlCAT2a was 1950bp in length, which encodes 650 amino acids, and SlCAT2b was 1635bp in length, which encodes 545 amino acids. SlCAT2a protein contained 14 putative transmembrane domains and SlCAT2b protein contained 12 putative transmembrane domains in silico analysis. SlCAT2a and