西方蜜蜂Lozenge基因的克隆鉴定及时空表达分析

Cloning and spatio-temporal expression of the gene Lozenge in Apis mellifera

Lozenge蛋白(Lz蛋白)是昆虫的重要转录因子,在昆虫胚胎发育过程中发挥重要作用。为研究Lozenge在西方蜜蜂Apis mellifera中的作用,本研究克隆了Lozenge基因,并对其进行生物信息学分析,同时基于荧光定量PCR技术检测该基因在西方蜜蜂不同发育时期(卵期、幼虫期、蛹期和成年蜂)和10日龄哺育蜂各组织的表达谱。生物信息学分析结果显示,Lozenge基因的开放阅读框(ORF)为1 554 bp,共编码517个氨基酸,预测分子量为54.63918 kDa,等电点为6.08;结构域预测分析发现Lozenge蛋白含有一个Runt结构域,多物种蛋白序列对比发现该蛋白同源性高。时期表达谱表明,该基因在第1日卵和第2日卵的表达量远高于其他时期,在卵期表达量随时间依次递减,幼虫期表达量极低,蛹期表达量呈先增后减的趋势,而成年蜂中均有表达;组织表达谱显示,该基因在哺育蜂头部、上颚腺中的表达量较高,而在腹部的表达量低。这些结果表明,Lozenge基因可能在西方蜜蜂胚胎期细胞发育过程、哺育蜂蜂王浆合成和分泌过程中发挥重要作用,这些结果为该基因功能的深入研究提供了重要的理论参考。

Lozenge protein,an important transcription factor in insect,plays a key role in the embryonic development.In order to study the role of LZ in honeybee,a Lozenge gene was cloned from Apis mellifera and analyzed using Bioinformatics.At the same time,expression profile of Lozenge gene was detected in different developmental stages(egg,larva,pupa and adult)and variety tissues of 10-day-old nurses via fluorescence quantitative PCR.Bioinformatics analysis showed that the open reading frame(ORF)of the gene was 1554 bp,encoding 517 amino acids;predicted molecular weight and isoelectric point was 54.63918 kDa and 6.08,respectively.It was found that Lozenge contained a Runt domain.The stage expression profile showed that the expression of the gene in the 1 d and 2 d egg stage was much higher than that in other stages.Its expression in the egg stage was decreased gradually with time,the expression in the larval stage was very low,the expression in the pupal stage was increased firstly and then decreased and it was expressed in adult stage.The tissue expression profile showed that the expression of the gene was higher in the head and mandibular gland,but lower in the abdomen.These results suggest that Lozenge gene may play an important role in the embryogenesis of A.mellifera,and may be involved in the nursing behavior of A.mellifera.The results provide an important theoretical reference for the further study of the function of this gene.