Wang Gaofeng, Peng Deliang, Sun Jianhua, Huang Wenkun, Peng Huan, Long Haibo
College of Life Sciences, Tianjin Normal University, Tianjin 300387, China.
Sheng Wu Gong Cheng Xue Bao. 2011 Jan;27(1):60-8.
The Cathepsin L-like cysteine proteinase genes (cpls) are multifunction genes related to the parasitic abilities of plant parasitic nematodes. A new cathepsin L-like cysteine proteinase gene (Dd-cpl-1) (GenBank Accession GQ 180107) was cloned from Ditylenchus destructor by RT-PCR and RACE. The cDNA sequence consisted of a 1 131 bp open reading frame (ORF) encoding 376 amino acid residues that were franked by a 29 bp 5'-untranslated region (UTR) and a 159 bp 3'-UTR. Genomic sequence analysis showed that Dd-cpl-1 contained 7 introns, obeyed the GT/AG rule in the splice-site junctions. Homology analysis showed that the identity was 77% between Dd-cpl-1 deduced protein Dd-CPL-1 and cathepsin L-like cysteine proteinase of Bursaphelenchus xylophilus. Multi-sequence alignment indicated that there were the catalytic triad (Cys183, His322 and Asn343) and two motifs ERFNIN motif and GNFD motif in deduced protein Dd-CPL-1. Cysteine proteinases phylogenetic analysis showed that Dd-cpl-1 belonged to the sub-clade of cathepsin L-like cysteine proteinases.
组织蛋白酶L样半胱氨酸蛋白酶基因(cpls)是与植物寄生线虫寄生能力相关的多功能基因。通过RT-PCR和RACE技术从腐烂茎线虫中克隆到一个新的组织蛋白酶L样半胱氨酸蛋白酶基因(Dd-cpl-1)(GenBank登录号:GQ 180107)。该cDNA序列由一个1131 bp的开放阅读框(ORF)组成,编码376个氨基酸残基,其5'端有一个29 bp的非翻译区(UTR),3'端有一个159 bp的UTR。基因组序列分析表明,Dd-cpl-1含有7个内含子,在剪接位点遵循GT/AG规则。同源性分析显示,Dd-cpl-1推导的蛋白Dd-CPL-1与松材线虫组织蛋白酶L样半胱氨酸蛋白酶的同一性为77%。多序列比对表明,推导的蛋白Dd-CPL-1中存在催化三联体(Cys183、His322和Asn343)以及两个基序ERFNIN基序和GNFD基序。半胱氨酸蛋白酶系统发育分析表明,Dd-cpl-1属于组织蛋白酶L样半胱氨酸蛋白酶亚分支。