摘要
【目的】克隆Q型烟粉虱(Bemisia tabaci)化学感受蛋白基因,分析其序列特征和表达特性,为深入研究该类基因功能奠定基础。【方法】根据桃蚜化学感受蛋白OS-D2a(ABM5558)的氨基酸序列,借助生物信息学搜索烟粉虱EST序列拼接了1个烟粉虱化学感受蛋白基因(BtabCSP1),以烟粉虱mRNA为模板,采用RT-PCR扩增克隆烟粉虱化学感受蛋白cDNA。采用半定量RT-PCR对不同发育阶段的烟粉虱样品中BtabCSP1的表达模式进行研究。【结果】BtabCSP1完整阅读框全长为2 626 bp,包含2个外显子和1个内含子,编码131个氨基酸残基,其中含有1个CX6CX18CX2结构域,为化学感受蛋白的典型特征。BtabCSP1在供试样品中均有表达,并且在2龄、3龄和伪蛹阶段表达量更高。聚类分析结果表明,烟粉虱与果蝇、桃蚜等物种遗传关系较远,化学感受蛋白基因在物种进化过程中较为活跃。【结论】从Q型烟粉虱若虫中克隆了1个化学感受蛋白基因BtabCSP1,该基因在烟粉虱不同发育阶段中均能表达,推测其对烟粉虱的生长发育具有重要的调控作用。
[Objective] The objective of this study is to clone the full length cDNA of the gene encoding chemosensory proteins from Bemisia tabaci Q biotype, and to analyze its expression pattern at different developmental stages. [Method] Based on the amino acid sequence of aphids （Myzus persicae） chemosensory proteins OS-D2a （ABM5558） reported previously, a pair of primers were designed based on simulation of gene splicing and RT-PCR was used to clone the chemosensory protein gene BtabCSP1. [Result] Sequencing and structural analysis showed that BtabCSP1 contains a 2 626 bp long complete reading frame （with 2 exons and 1 intron） and encodes 131 amino acid residues with a CX6CX18CX2 domain, the typical characteristics of a ehemosensory protein gene. As displayed by semi-quantitative RT-PCR amplification from total RNA ofB. tabaci of various stages, BtabCSP1 was expressed in all samples and the expressions were more apparent in the 2nd instar, 3rd instar and 4th instar （pupal stage） nymphs. The amino acid （encoded by BtabCSP1） sequence cluster analysis showed that B. tabaci was distantly related to fruit flies and aphids and chemosensory protein gene was active in the process of species evolution. [Conclusion] BtabCSP1 was isolated from B. tabaci Q biotype, and the expression pattern suggested the close relationship between the expression of this gene and the development ofB. tabaci.
出处
《中国农业科学》
CAS
CSCD
北大核心
2012年第18期3892-3898,共7页
Scientia Agricultura Sinica
基金
国家转基因新品种培育重大专项项目(2009ZX08012-007B)
