The Cloning Of TFL1 And SOC1 Genes Of Rose And The Construction Of Expression Vector

Chinese rose is one of the four cut flowers, beautiful flower, smell quietly elegant. By the use of technique, human intervention to Chinese rose flower, can obtain higher economic value, medicinal value and ornamental value. Blossom pathways including pHotoperiod pathway flowers, autonomous pathway and gibberellin four ways, they interact with each other, mutual cross link, work together to plant flowering regulation. This experiment successfully extracted the RNA of Chinese rose which grow up from Shanxi agricultural university, and cloned the two regulating the flowering gene,TFL1 and SOC1. Using bioinformatics analysis technique, the gene sequences, the amino acid sequence, and the secondary and the tertiary structure of protein were analysed. The study also built gene expression vector of TFL1 and SOC1.1 The total RNA of Chinese rose which grow up from shanxi agricultural university was extracted.cDNA was synthesised by reverse transcription. Using Premier & Oligo software, we designed TFLl gene specific primers, upstream Primer:TFL1f3 ’5’-GCTCAAGGAA AACAGAAAAGGAT-and downstream primers:TFL1r 5’-CAGAAGCATCACTTGGT TGTTTATC-3’. Using PCR technique, TFL1 of Chinese rose flower of Shanxi agricultural university was amplified, size of 899 bp. Using Premier & Oligo software, we designed SOCl gene specific primers, upstream Primer:SOC1 51-GTAGATTGCTTTTGAGAACCC ATTTG-3’and downstream primers:SOC1 5’-CAGAACATACGCAAAACAATCGGC -3’. Using PCR technique, SOC1 of Chinese rose flower of Shanxi agricultural university was amplified, size of 725 bp.2 Through bioinformatics analysis, TFL1 of Chinese rose flowering of Shanxi agricultural university had the homologous of 99% with the genes of Rosa hybrid cultivar RhTFLl (TFL1) (landing JF806626.1) from NCBI, there were three mutated bases.By simulating the structure of the protein, secondary structure analysis showed that the alpHa helix, the two folded beta had subtle differences; tertiary structure was no significant difference.SOC1 of Chinese rose flowering of Shanxi agricultural university had the homologous of 99% with the genes of Rosa hybrid cultivar SOCl-like protein (SOC1) (login KC121576.1) from NCBI, there were three mutated bases.By simulating the structure of the protein, secondary structure analysis showed that the alpHa helix, the two folded beta had subtle differences; tertiary structure was significant difference. Protein folding and space conformation determines the function of protein, the three mutations located in the core of SOCl. SOCl gene is an important member of plant flowering integron genes. It follows that the shanxi agricultural university, Chinese rose flowering change may caused by SOCl gene mutations.3 TA plasmid was identified by PCR test. The pCAMBIA2300-TFL1; vector was built by Kpn I and Sal I double enzymic digestion; The PMV-SOC1 vector was built by Kpn I and Xho I double enzymic digestion.