中华蜜蜂离子型受体AcerIR76b、AcerIR75f.1生物信息学和差异表达分析

Differential Expression and Bioinformatic Analysis of Ionotropic Receptor Gene AcerIR76b and AcerIR75f.1 from Apis cerana cerana

【目的】从中华蜜蜂Apis cerana cerana触角转录组测序结果中筛选获得中华蜜蜂离子型受体IR76b、IR75f.1的基因序列,对其进行蛋白结构预测和表达谱分析,为该基因的功能研究提供依据。【方法】利用多种生物信息学软件对其编码蛋白的理化特性进行分析;采用qRT-PCR技术对AcerIR76b、AcerIR75f.1在中蜂1日龄工蜂和采集蜂、1日龄雄蜂和性成熟雄蜂各部位(触角、头、胸、腹和足)中mRNA的表达情况进行比较分析。【结果】成功获得了AcerIR76b、AcerIR75f.1的完整开放阅读框ORF序列,全长分别为1635 bp和1686 bp,分别编码545、562个氨基酸。预测AcerIR76b分子量为63.09 ku,AcerIR75f.1分子量为62.28 ku,它们均具有3个跨膜结构。qRT-PCR结果显示,AcerIR76b在1日龄工蜂和采集蜂、1日龄雄蜂和性成熟雄蜂的各部位中均有表达,但在触角中的表达量极显著的高于其它部位(P<0.01);AcerIR75f.1在1日龄工蜂和采集蜂中,均在触角中高表达,而在1日龄雄蜂和性成熟雄蜂中,均在足部高表达。【结论】AcerIR76b和AcerIR75f.1的编码产物均具有昆虫离子型受体的典型结构特征,2个离子型受体在中华蜜蜂工蜂和雄蜂各个部位均有表达,推测其不仅参与气味分子的识别过程,在味觉感知中也同样发挥着一定作用。

[Objective]The gene sequence of ionotropic receptor IR76b and IR75f.1 was screened from the transcriptome sequencing results of Apis cerana cerana,and the protein structure prediction and expression profiling were analyzed so as to lay afoundation for the research on this gene,which provides a theoretical basis for further exploration of the physiological functions of these genes.[Method]The physicochemical properties of the encoded protein were analyzed by a variety of bioinformatics software.The method of qRT-PCR was used to detect the expression profiles of AcerIR76b and AcerIR75f.1 mRNA in five parts(antennae,head,thorax,abdomen and legs)of 1-day-old workers and foragers,1-day-old and sexually mature drones.[Result]The complete ORF of AcerIR76b and AcerIR75f.1 were successfully obtained,which were 1635 bp and 1686 bp in length and encoded 545 and 562 amino acids.The molecular weight was predicted to be 63.09 kDa about AcerIR76b,and 62.28 ku about AcerIR75f.1,they had three transmembrane structures.The results of qRT-PCR showed that AcerIR76b was expressed in all parts of the workers and drones,but the expression in the antennae was significantly higher than that in other parts(P<0.01).In 1-day-old workers and foragers,AcerIR75f.1 was highly expressed in the antennae,while in 1-day-old and sexually mature drones,this gene was highly expressed in the legs.[Conclusion]The coding products of AcerIR76b and AcerIR75f.1 owned typical characteristics of insect ionotropic receptors.Two ionotropic receptors were expressed in all parts of workers and drones,implying that they not only participated in the recognition process of odor molecules but also played a role in taste perception.