摘要
【目的】克隆淡色库蚊一个全新的氯菊酯抗性相关基因PR-XP1的全长序列,分析、预测其编码蛋白的结构与功能,为阐明该基因的抗性机制及研制新型卫生杀虫剂奠定基础。【方法】依据抗性与敏感品系库蚊差异表达的EST片段(GenBank号:EC093826.1),设计特异性PCR引物,采用RACE技术,克隆淡色库蚊氯菊酯抗性相关基因PR-XP1的全长cDNA序列,运用生物信息学软件对其同源性与系统进化进行分析,并对PR-XP1编码蛋白的结构与功能进行预测。【结果】获得了全长为2 004bp的淡色库蚊氯菊酯抗性相关基因PR-XP1,该基因具有完整的CDS区,编码249个氨基酸。同源性比对与系统进化分析表明,PR-XP1基因核苷酸序列与致倦库蚊(GenBank号:XM_001862469.1)、埃及伊蚊(GenBank号:XM_001658032.1)和冈比亚按蚊(GenBank号:BX021208.1)相关基因的同源性分别为95%,83%和70%;其编码蛋白的氨基酸序列与致倦库蚊、埃及伊蚊、西方蜜蜂、入侵红火蚁、佛罗里达弓背蚁、印度跳蚁等昆虫中"假设性蛋白"的氨基酸序列同源性均达46%以上。结构分析显示,PR-XP1基因编码蛋白可能为具有2个跨膜螺旋的膜蛋白,跨膜区形成了一个特异性的"U"形结构;该基因还具有1个微弱的信号肽位点和24个磷酸化位点。【结论】克隆得到了一个全新的淡色库蚊氯菊酯抗性相关的"保守性假设蛋白"基因PR-XP1,推测其功能与氯菊酯抗性相关。
【Objective】 The full length cDNA sequence of a new permethrin-resistance associated gene PR-XP1 of Culex pipiens pallens was obtained and its coding protein in bioinformatics was analyzed,which would lay foundation for clarifying the resistance mechanism of this gene and developmenting new pesticides.【Method】 Primers were designed according to EST（GenBank ID：EC093826.1） from the different expressions between permethrin-susceptible and-resistant strains of Culex quinquefasciatus,and they were used for cloning the full length cDNA sequence of gene PR-XP1 in RACE technique.After the homeology and phylogency were analyzed,the structure and function prediction of the protein encoded by gene PR-XP1 were performed in bioinformatic.【Result】 The length of cDNA sequence was 2 004 bp,which had a complete CDS region coding 249 amino acids.Homology comparisons and phylogenetic analysis showed that the nucleotide sequence of gene PR-XP1 of Culex pipiens pallens shared 95%,83% and 70% similarity with the gene in Culex quinquefasciatus（GenBank ID：XM_001862469.1）,Aedes aegypti（GenBank ID：XM_001658032.1）,and Anopheles gambiae（GenBank ID：BX021208.1）.And more than 46% similarity with Culex quinquefasciatus,Aedes aegypti,Apis mellifera,Solenopsis invicta,Camponotus floridanus,Harpegnathos saltator in amino acid.The bioinformatic analysis results showed that the protein coded by gene PR-XP1 was a membrane protein with two transmembrane helices,which were constituted a specific ＂U＂ structure.The gene had one weak signal peptide cleavage point and twenty-four phosphorylation sites.【Conclusion】 A new permethrin-resistance associated conserved hypothetical protein gene PR-XP1 was cloned in Culex pipiens pallens,which would play an important role in permethrin-resistance.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2011年第11期194-200,共7页
Journal of Northwest A&F University(Natural Science Edition)
基金
西北农林科技大学青年学术骨干研究基金项目(01140302)
关键词
淡色库蚊
氯菊酯抗性
保守性假设蛋白
生物信息学分析
Culex pipiens pallens
permethrin-resistance
conserved hypothetical protein
bioinformatic analysis
