| 摘要: |
| 【目的】克隆拟黑多刺蚁（Polyrhachis vicina Roger）5-HT2A受体基因的cDNA序列，并对其核苷酸序列和系统进化进行分析，检测其在拟黑多刺蚁不同发育阶段幼虫和不同品级成虫mRNA的表达情况，为探究5-HT2A受体基因对拟黑多刺蚁生长发育的调控规律提供证据。【方法】从拟黑多刺蚁中提取总RNA，RT-PCR扩增并克隆Pv5 HT2A受体基因；用ProtParam预测蛋白质序列，Blast比较蛋白质序列同源性，用Clustal X进行核酸和氨基酸序列比对，MEGA 6.0比邻法生成系统进化树。利用荧光实时定量PCR检测Pv5 HT2A受体基因在拟黑多刺蚁不同发育阶段幼虫及不同品级成虫mRNA的表达水平。【结果】克隆得到Pv5-HT2A受体基因的cDNA 序列，全长2 965 bp，开放阅读框长1 926 bp，编码641个氨基酸。疏水性分析显示,Pv5-HT2A受体氨基酸具有7个跨膜疏水区，属于典型的G蛋白偶联受体类型。系统发育分析表明，拟黑多刺蚁与红收获蚁的亲缘关系最近，相似度达69%。荧光实时定量PCR结果显示，拟黑多刺蚁5-HT2A mRNA在不同发育阶段和不同品级成虫中均有表达,其中卵、3龄幼虫及蛹期的表达量较高;成虫中,雄蚁表达量最高，雌蚁的表达量最低。【结论】拟黑多刺蚁与其他昆虫的5 HT2A受体基因存在相关性，推测5 HT2A受体基因对拟黑多刺蚁的生长发育具有一定调节作用。 |
| 关键词: 拟黑多刺蚁 5-HT2A受体基因 克隆 发育调控 基因表达 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金面上项目（31571276，31171195) |
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| Cloning and expression analysis of 5-HT2A gene in Polyrhachis vicina Roger (Hymenoptera:Formicidae) |
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YANG Sen,TAN E,XI Gengsi
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| Abstract: |
| 【Objective】Polyrhachis vicina Roger Pv5-HT2A gene was cloned,its nucleotide sequence and system evolution were analyzed,and its expressions in different developmental stages were investigated in this paper to provide evidence for the participation of Pv5-HT2A receptor in modulation of P.vicina development and further exploration of Pv5-HT2A functions.【Method】The total RNA was extracted from P.vicina,the Pv5-HT2A receptor gene was amplified and cloned,and the protein sequence was predicted by ProtParam.The homology was compared by Blast,nucleic acids and amino acid sequence alignment were conducted by Clustal X and phylogenetic tree was generated by MEGA 6.0 nearest neighbor method.The Pv5-HT2A receptor mRNA expressions were detected in different developmental stages and castes using real-time PCR.【Result】The obtained full length cDNA of Pv5-HT2A receptor gene was 2 965 bp.The open reading frame (ORF) of 1 926 bp encoded 641 amino acids.Hydrophobic analysis showed that the Pv5-HT2A receptor had seven transmembrane hydrophobic regions.The phylogenetic tree analysis revealed that P.vicina had close relationship with Pogonomyrmex barbatus.The real-time PCR results showed that Pv5-HT2A mRNA was detected at all tested development stages.Pv5 HT2A mRNA was expressed highly in embryo,3rd larvae and pupae.The Pv5-HT2A expression level in male was higher than in female.【Conclusion】The 5-HT2A receptor of P.vicina was cloned successfully and it had correlation with the genes of other insects.It is speculated that 5 HT2A receptor gene plays certain role in growth and development of P.vicina. |
| Key words: Polyrhachis vicina Roger 5-HT2A receptor gene clone development regulation gene expression |
