Cloning Of Sucrose Phosphate Synthase Gene In Tomato Fruit And Study On Tomato Genetic Transformation System

In this research, we cloned sucrose phosphate synthase gene complete cDNA from tomato fruit via RT-PCR, analysed its sequence. Its access number in GenBank is AY726439. The regeneration system of "T6"tomato was optimized. Melon fruit acid invertase gene antisense vector was transferred into"T6"tomato mediated by agrobacterium tumefaciens strain LBA4404. Kanamycin-resistance tomato plantlets were obtained. The details are as follows: "20-19"tomato fruit total RNA was extracted via one step of Thiocynate-Phenol-Chloroform method. The result showed that the RNA extracted had high purity , met the need of experiment. The RT-PCR primers were designed based on the consensus domain of sucrose phosphate synthase gene such as tomato and potato registered in GenBank. The aimed product was amplified from the total RNA isolated from "20-19"tomato fruit via RT-PCR. Then it was inserted into T-vector and sequenced. The result showed that this fragment was the aimed gene complete cDNA. It contained 3373bp, with a open-reading-frame of 3162bp, and 1054 amino acids determined by this fragment. The result of sequence analysis showed that it shared 98% homology in nucleotide sequence with tomato SPS gene, whose access number is AB051216 in GenBank. The result of comparison with several other species showed that it shared 96% homology in nucleotide sequence with potato SPS gene, whose access number is X73477; 92% homology with tobacco SPS gene whose access number is AF194022; 82% homology with sweet potato SPS gene whose access number is AF439861. This gene had been registered in...