中华蜜蜂GnRH相关受体的基因克隆及表达研究

摘要

[目的]克隆中华蜜蜂（Apis cerana cerana）促性腺激素释放激素（gonadotrophin releasing hormone，GnRH）相关受体基因，对其进行表达研究。[方法]利用RT—PCR技术克隆了中华蜜蜂的GnRH相关受体基因的cDNA序列，并对其进行生物信息学分析和表达产物的原位杂交组织化学研究。[结果]序列分析显示，GnRH相关受体的cDNA序列全长1177bp，开放阅读框长1050bp，编码349个氨基酸残基。推导的氨基酸序列具有7个跨膜区；预测的分子量和等电点分别为40．6kD和9．54。聚类分析显示该蛋白质与其他已知昆虫的GnRHRⅡ具有较近的亲缘关系。原位杂交显示GnRH相关受体基因在中华蜜蜂的脑、精巢、脂肪体以及肠道等组织均有表达。[结论]该研究结果表明GnRH相关受体为昆虫提供了生殖和新陈代谢之间的分子纽带，其在协调昆虫的生殖活动和环境状况的关系中执行了一定的功能。%[Objective] This study was to clone the GnRH （gonadotropin-releasing hormone）, and to investigate its expression in Apis cerana cerana. [Method] The cDNA sequence of GnRHR gene was amplified from Apis cerana cerana by using RT-PCR techniques. It was conducted with bioinformatics analysis and the in situ hybridization histochemistry of its expression products was studied. [Result] The sequence analy- sis showed that the full cDNA sequence was 1 050 bp with the open reading frame of 1 050 bp, and it encoded 349 amino acid residues. The deduced amino sequence included 7 transmembrane regions, and the predicted molecular mass and isoelectric point were 40.6 kD and 9.54, respectively. The cluster analysis showed that the GnRHR from ＇,4. cerana cerana had close relationship to the GnRHR II from other insects. In situ hybridization showed that Bee-GnRHR staining was specifically localized to the brain, intestine, fat body and testis. [Conclusion] The results indicated that the GnRHR provided molecular bond for the reproduction and metabolism for insects, and suggested a functional role for bee-GnRHR signaling in the coupling of reproduction activities and environment conditions.