| 摘要: |
| 【目的】扶桑绵粉蚧是我国重要的入侵生物，其繁殖能力强，能危害棉花、扶桑、向日葵、南瓜、番茄等多种我国重要的经济作物。克隆扶桑绵粉蚧谷胱甘肽S-转移酶（GSTs）基因,可为揭示扶桑绵粉蚧GSTs的生理功能提供参考。【方法】利用RT-PCR和RACE技术克隆扶桑绵粉蚧谷胱甘肽S-转移酶基因全长，并采用生物信息学方法分析其结构特征，用实时荧光定量PCR的方法研究其各个虫态的表达谱。【结果】克隆了扶桑绵粉蚧谷胱甘肽S-转移酶基因全长序列,该基因的开放阅读框包含651 bp的片段，编码217个氨基酸。DNA编码区由4个内含子和5个外显子组成,内含子的长度分别为90，123，67和70 bp；分隔的5个外显子的长度分别为18，50，96，80和500 bp。功能域分析结果显示，该蛋白在N末端和C末端均有GST的类似结构位点。多序列比对及系统进化树构建结果表明，该蛋白属Zeta家族GSTs，将其命名为PsGSTzl。PsGSTzl mRNA在扶桑绵粉蚧的不同虫态中都有表达，在1龄幼虫中的表达量最高。【结论】成功克隆的PsGSTzl基因在不同虫态中差异表达，为进一步揭示该基因在虫体的代谢作用奠定了基础。 |
| 关键词: 扶桑绵粉蚧 谷胱甘肽S-转移酶 克隆 表达谱 |
| DOI： |
| 分类号: |
| 基金项目:农业部公益性行业科研专项（201103026）；仲恺农业工程学院科研基金项目（G3100004） |
|
| Molecular cloning and expression of different developmental stages of glutathione S-transfeses gene in mealybug Phenacoccus solenopsis Tinsley (Hemiptera:Pseudococcidae) |
|
|
| Abstract: |
| 【Objective】Phenacoccus solenopsis was one of the most important pests in our country,which can damage cotton,hibiscus,sunflower,pumpkin,tomatoes and other important economic crops.The research of glutathione S-transferase gene provided a foundation for revealing the GSTs physiological function.【Method】The RACE method was used to obtain the full-length gene,bioinformatics methods to analyze the structural features,the real-time PCR method to study the expression profiles of the various instars.【Result】We cloned the full length of glutathione S-transferase gene of P.solenopsis Tinsley.The gene contained an open reading frame fragment of 651 bp,encoding 217 amino acids.DNA coding region was composed of four introns and five exons.Introns length was 90,123,67 and 70 bp,while the length of the separated four exons was 18,50,96,80 and 500 bp.Functional domain analysis revealed that the protein in both N-terminal and C-terminal got the GST-like structure sites.Multiple sequence alignment and phylogenetic tree construction revealed that this gene was a protein family of insect-specific Zeta GSTs which we named as PsGSTzl.PsGSTzl mRNA in P.solenopsis expressed in different instars,while the first larvae was one of the highest expressions.【Conclusion】The successfully cloned P.solenopsis glutathione S-transferase gene and the expression in different instars were different,which could further reveal the role of genes in the parasite’s metabolic basis. |
| Key words: Phenacoccus solenopsis glutathione S-transfeses cloning expression profile |
