拟黑多刺蚁Broad-Complex基因的分子克隆及表达分析  

Molecular cloning and expression analysis of the Broad-Complex gene in Polyrhachis vicina Roger

作　　者：张娟娟 奚耕思[1] 赵玉峰 岳秀敏 ZHANG Juanjuan;XI Gengsi;ZHAO Yufeng;YUE Xiumin(School of Life Sciences,Shaanxi Normal University,Xi′an 710119,Shaanxi,China)

机构地区：[1]陕西师范大学生命科学学院,陕西西安710119

出　　处：《陕西师范大学学报（自然科学版）》2020年第5期56-64,77,共10页Journal of Shaanxi Normal University：Natural Science Edition

基　　金：国家自然科学基金(31571276,31171195)。

摘　　要：采用RT-PCR(revese transcription-polymerase chain reaction)、RACE(rapid amplification of cDNA ends)等技术,依据转录组测序结果,从拟黑多刺蚁中克隆得到与Broad-Complex(BR-C)同源的全长cDNA序列,命名为PvBR-C(登录号为MF596490.1);采用生物信息学方法对其cDNA和编码蛋白质的理化性质、蛋白质二级及三级结构、分子系统进化关系等进行预测和推断。PvBR-C基因的cDNA全长3061 bp,包含1个1302 bp的开放阅读框,编码由433个氨基酸组成的蛋白质。生物信息学分析表明,PvBR-C与BR-C蛋白具有高度同源性,且具有BTB和Zf-H2C2结构域。RT-qPCR(real-time quantitative polymerase chain reaction)分析表明,PvBR-C在拟黑多刺蚁不同发育阶段和不同成虫品级中的表达水平差异较大,幼虫发育时期表达量最高;在不同品级成体的不同部位均有表达,但在生殖蚁的腹部和工蚁的头部高表达。由上述实验结果推测PvBR-C可能对于拟黑多刺蚁不同发育阶段虫体及不同品级成虫的调节具有特异性,对调节生殖蚁的生殖和消化功能以及工蚁的行为发挥着重要作用。In this study,the full-length cDNA of a BR-C homolog was obtained from the Polyrhachis vicina Roger based on the result of RNA-Seq using RT-PCR and rapid amplification of cDNA ends techniques.The homolog was designated as PvBR-C(GenBank accession no.MF596490.1).The nucleotide sequence and deduced amino acid sequence of PvBR-C were analyzed and characterized by bioinformatic methods.The results showed that the full-length cDNA of PvBR-C was 3061 bp,including a 1302 bp open reading frame,and encoding a precursor protein of 433 amino acid residues.The PvBR-C exhibited high homology with BR-C protein and contained the BTB and Zf-H2C2 domains.Real-time quantitative polymerase chain reaction(RT-qPCR)analysis revealed that the PvBR-C expression level varied significantly in different development stages and adult castes.The expression was highest in the larral stage.It was expressed in all studied adult tissues with high expression levels detected in the abdomen of reproductive ants and the head of the worker ants.Our data demonstrated that PvBR-C may affect the regulation of different developmental stages and different grades of Polyrhachis vicina Roger,and may play important roles in regulating the reproductive and digestive functions of reproductive ants and the behavior of worker ants.

关 键 词：拟黑多刺蚁 BROAD-COMPLEX CDNA RT-QPCR 表达分析 

分 类 号：TP391.42[自动化与计算机技术—计算机应用技术]