草原樱桃PGIP基因的克隆及生物信息学分析

Cloning and Bioinformatic Analysis of PGIPGene fromPrunus caoyuan

为了克隆草原樱桃多聚半乳糖醛酸酶抑制蛋白基因,并进行生物信息学分析。以草原樱桃叶片基因组为模板,PGIP基因保守序列设计引物,通过PCR扩增获得约1 kb的DNA片段。序列分析表明,该基因全长1 193 bp,包含有1个完整的开放阅读框,由2个外显子和1个内含子构成,外显子总长990 bp,编码330个氨基酸,其编码的氨基酸序列中含有一段典型的亮氨酸重复序列,GenBank登录号为GU068977;该基因与中国李、杏、桃、马哈利樱桃、梅等李属植物的PGIP基因序列一致度达95%～99%。系统进化分析显示,属内亲缘关系较近、属间亲缘关系较远的特点。克隆了草原樱桃PGIP基因,为樱桃抗病育种提供一条新的基因资源。

Disease resistance mechanism was studied by methods of cloning of the PGIP gene in Prunus caoyuan.A DNA fragment about 1 kb was amplified from the genomic DNA of Prunus caoyuan leaves by PCR with a pair of specific primers based on the conserved sequences of the PGIP genes of genus Prunus.Sequence analysis showed that the fragment contains a full coding region of 1 193 bp（GenBank accession：GU068977）.This sequence had a full open reading frame encoding the PGIP,and eontained two exons interrupted by one intron.The total exons were comprised by 990 bp of deoxynucleotide encoding 330 amino acid.A conserved leucine-rich fragmenthad existed in the derived protein sequence.Sequencing analysis showed that it was 95% to 99% identical with the sequences of Prunus PGIP genes including P.salicina,P.armeniaca,P.persica,P.mahaleb,P.mume.Phylogenic tree showed that genetic relationship within the genus was closer and between the genera was farther.A PGIP gene of Prunus caoyuan was cloned.As a result,a gene resource was provided for molecular breeding of plants.