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[来自毁灭茎线虫的一个新的组织蛋白酶L样半胱氨酸蛋白酶基因的克隆与序列分析]

[Cloning and sequence analysis of a new cathepsin L-like cysteine proteinase gene from Ditylenchus destructor].

作者信息

Wang Gaofeng, Peng Deliang, Sun Jianhua, Huang Wenkun, Peng Huan, Long Haibo

机构信息

College of Life Sciences, Tianjin Normal University, Tianjin 300387, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2011 Jan;27(1):60-8.

Abstract

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.

摘要

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。

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