中矮1号梨砧木(S_2)PGIP基因的克隆及序列分析  

Cloning and sequence analysis of PGIP gene from pear dwarf stock S_2 (Pyrus ussuriensis)

作　　者：蒋豪[1] 汤浩茹[1] 古英洪[1] 张勇[1] 罗娅[1] 

机构地区：[1]四川农业大学林学园艺学院,四川雅安625014

出　　处：《果树学报》2008年第4期462-466,共5页Journal of Fruit Science

基　　金：国家自然科学基金(编号:30671454);教育部"新世纪优秀人才支持计划"(编号:NCET-04-0905);高等学校全国百篇优秀博士学位论文作者专项(编号:200253)

摘　　要：利用RT-PCR技术,根据GenBank中梨属PGIP基因序列设计1对特异引物,以梨矮化砧木S2叶片总RNA为模板,克隆到1条约1100bp的cDNA片段,将其与pMD18-Tvector连接后转化Escherichia coli JM109,对筛选到的阳性克隆进行序列测定并使用生物信息学方法对所得结果进行综合分析。结果表明,克隆片段为梨PGIP基因,该cDNA编码330个氨基酸,预测分子量为36388ku,第1～24个氨基酸残基是信号肽。与梨属其它种中已获得的PGIP核苷酸序列开放阅读框(Open reading frame,ORF)的同源性为97.5%～99.6%,氨基酸序列的同源性为97.6%～99.1%。A 1 038 bp sequence of polygalacturonase-inhibiting protein （PGIP） gene （Genbank Accession Number： EU107274） was obtained from Pyrus ussuriensis by RT-PCR using a pair of specific primers. PCR product was cloned into pMD18-T vector and then transformed into Escherichia coli JM109. Positive clones were picked out and sequenced. The results revealed that PuPGIP encodes a 330 amino acid protein with a predicted molecular weight of 36 388 ku. The predicted signal peptide was formed by 24 residues from 1 to 24. The cloned cDNA exhibits a homology of the open reading frame （ORF） of the nucleotide and deduced amino acid sequences of 97.5% to 99.6% and 97.6% to 99.1%, respectively.

关 键 词：梨(Pyres ussuriensis) 矮化砧木 基因克隆 PGIP基因 序列分析 

分 类 号：S661.2[农业科学—果树学]