Cloning and Bioinformatics Analysis of Gene Sequence Encoding ACC Synthase from Paeonia suffruticosa

ACC synthase is the key enzyme in ethylene biosynthsis.Based on reported cDNA sequence(DQ337250) encoding ACC synthase from Paeonia suffruticosa Andr.,a pair of specific PCR primers was designed.With the total DNA extracted from fresh leaves of Paeonia suffruticosa cv.Luoyanghong as DNA template,PCR products about 2 200 bp in length were successfully amplified by high-fidelity KOD-Plus DNA polymerase.Then the PCR products were cloned to pMD18-T vector.The sequencing result showed that the full-length of the genomic sequence of ACSisolated from Paeonia suffruticosa Andr.was 2 251 bp containing 4 extrons and 3 introns,and all the splicing sites were conservative at 5′ donor with GU and 3′ receptor with AG.The 4 extrons were located from 1 to 174,267 to 398,483 to 643 and 1 240 to 2 251 bp,respectively.The sequence information has been submitted to GenBank with the accession number of FJ769773.The ACS protein contained 492 amino acids and its similarities to other ACS in Malus×domestica,Eriobotrya japonica,Pyrus communis and Pelargonium×hortorum were up to 74%-76%.Bioinformatic analysis revealed that the molecular weight and pI of ACS was 54.9 kD and 6.8,respectively.It also showed that ACS was an unstable,non-secretory hydrophilic protein,being located in cytoplasm.