摘要： 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。
关键词:
草原樱桃,
PGIP基因,
基因克隆,
生物信息学
Abstract: Disease resistance mechanism was studied by methods of cloning of the PGIPgene in Prunus caoyuan． A DNA fragment about 1 kb was amplified from the genomic DNA of Prunus caoyuanleaves by PCR with a pair of specific primers based on the conserved sequences of the PGIPgenes 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 PGIPgenes 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 PGIPgene of Prunus caoyuanwas cloned． As a result，a gene resource was provided for molecular breeding of plants．
Key words:
Prunus caoyuan,
PGIPgene,
Gene cloning,
Bioinformatic
中图分类号:
于文全, 刘海荣, 杨晓华, 赵恒田. 草原樱桃PGIP基因的克隆及生物信息学分析[J]. 华北农学报, 2012, 27(6): 38-42. doi: 10.3969/j.issn.1000-7091.2012.06.009.
YU Wen-quan, LIU Hai-rong, YANG Xiao-hua, ZHAO Heng-tian. Cloning and Bioinformatic Analysis of PGIPGene from Prunus caoyuan[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(6): 38-42. doi: 10.3969/j.issn.1000-7091.2012.06.009.