Cloning of Partial Sequence of GPAT Gene in Citrus and Its SNP Analysis

GPAT gene is an important gene with cold resistance of plants.A pair of degenergte primers were designed according to the conserve regions of GPAT gene of many kinds of plants reported in Genebank.Eight cDNA documents of GPAT gene in Citrus maxima cv.Guanximiyou,Citrus maxima cv.Majiayou,Citrus mangshanensis,Citrus limon,Citrus sinensis.Xingguotiancheng,Citrus maxima cv.Xinmuyou,Poncirus trifoliate and Citrus maxima cv.Lingnanshatianyou were cloned by using RT-PCR technology.The biology program of DNAStar5.0 was used to analyze the eight document sequences.The best nucleotide similarity was 97.8% among the eight gene documents and the similarity was above 72.5% with other plants'GPAT gene.Every nucleotide sequence encoded 105 amino acids according to derivation.The best amino acid similarity was 95.2% among the eight gene documents and the similarity was above 68.9% with other plants'GPAT gene.Fourteen variational sites of single nucleotides were discovered in the eight gene documents.Six amino acid sites transformed because of the variation of fourteen single nucleotides.Accoring to the calculation result,the polymorphism frequence was 1SNP/22.5 bp,the nucleotide and amino acid degree of the variation of nucleotide and amino acids were 4.45% and 5.71%.The genetic diversity of cloning documents was analyzed by using the clustering procedure of NJ(Neighbor-Joining).