Cloning and Sequence Analysis of ACC Oxidase Gene from Peach

Ethylene regulation of fruit ripening and senescence is an endogenous hormone.Therefore,regulating the biosynthesis of ethylene effectivly is the key to controlling fruit ripening.1-Aminocyclopropane-1-Carboxylic acid oxidase(ACO)gene is one of the rate-limiting enzyme in ethylene biosynthesis of plant.So total RNA was extracted from mature Baifen peach in this study,and clone ACO gene.The use of antisense technology is to inhibit the formation of ethylene,delay fruit ripening and improve the hardness of peach.The cloned fragment connected with pMD18-T vector and transformed into E.coli DH5α with PCR,restriction enzyme digestion and sequencing.Sequence analysis showed that the gene had the length of 1 246 bp,which contained a 960 bp coding region,encoded a total of 319 amino acids,and compared with nucleotide sequence and amino acid sequence homology of ACC oxidase gene(AF319166)landed were 99% and 98%,which showed the successful cloning of the ACC oxidase gene.With the bioinformatics analysis,the predicted ACC oxidase protein's molecular formula was C1626H2542N420O485S12,whose relative molecular mass was 36.12 kDa and isoelectric point was 5.27;It is predicted that this protein was hydrophilicity and non-secretory protein,and there was no signal peptide.Characteristic prediction results indicated that the secondary structure of ACC oxidase protein mainly included α-helix,β-sheet and random coil,and this protein belonged to a kind of a compact globular domain.