摘要
旨在明确中华蜜蜂Smoothened基因(AcerSmo)的序列特征及表达特性,为进一步研究其在蜜蜂体内的功能奠定理论基础,对中华蜜蜂AcerSmo基因DNA序列进行扩增鉴定,运用多种生物信息学软件对其进行结构和功能分析,并采用RT-qRCR对中华蜜蜂不同组织和发育时期的表达特性进行检测。序列分析结果表明,基因cDNA全长3 495 bp(GenBank登录号:OP616035),其中开放阅读框(ORF)区域2 952 bp,共编码936个氨基酸。该蛋白理论分子质量为111.15 ku,理论等电点为8.82,属于不稳定的两性蛋白。该蛋白N-末端有一段含21个氨基酸的信号肽,有6个跨膜结构域,亚细胞定位主要在内质网和细胞核。该蛋白二级结构和三级结构以无规则卷曲(50.76%)为主,其次为α-螺旋(25.33%),从昆虫到哺乳动物,AcerSmo蛋白序列高度保守;荧光定量PCR结果表明,AcerSmo在30日龄中华蜜蜂不同组织中均存在表达,而在触角与头的表达量显著高于其他组织,在中华蜜蜂触角与头组织中,5日龄时AcerSmo的mRNA表达量最高。AcerSmo蛋白具有Smoothened蛋白的保守结构,且在触角中呈高丰度表达,推测AcerSmo基因可能在蜜蜂的嗅觉识别过程发挥着重要作用。
The aim of this study was to clarify the sequence characteristics and expression properties of the Smoothened gene in Apis cerana cerana.The DNA sequence of AcerSmo was amplified and identified,its structure and function were analysed using various bioinformatics software,and the expression characteristics were examined by RT-qPCR for different tissues and developmental periods of Apis cerana cerana.Sequence analysis showed that the AcerSmo was 3495 bp(GenBank accession number:OP616035),the open reading frame(ORF)region was 2952 bp,encoding 936 amino acids.The molecular weight of this protein was 111.15 ku and the predicted isoelectric point was 8.82,which had a signal peptide containing 21 amino acids at the N-terminus,and there were 6 transmembrane structures domains.Subcellular localization mostly takes place in the nucleus and endoplasmic reticulum.The secondary and tertiary structures of this protein were dominated by irregular coiled(50.76%),followed byα-helix(25.33%).The sequence of AcerSmo protein sequence was highly conserved from insects to mammals.The results of RT-qPCR showed that AcerSmo was expressed in different tissues of Apis cerana cerana at the age of 30 days and the expression level of AcerSmo in the antennae and head were significantly higher than in other tissues,and in the tissues of the antennae and head of the Apis cerana cerana,the mRNA expression of AcerSmo was the highest at 5 days of age.AcerSmo has the conserved structure of Smoothened protein and was highly expressed in high abundance in antennae,which may play an important role in the olfactory recognition process of honey bees.
作者
于点点
申红英
王珏
郭媛
郭丽娜
YU Diandian;SHEN Hongying;WANG Jue;GUO Yuan;GUO Lina(College of Animal Science,Shanxi Agricultural University,Taigu 030801,China;College of Horticulture,Shanxi Agricultural University,Taigui 030801,China)
出处
《华北农学报》
CSCD
北大核心
2024年第3期230-238,共9页
Acta Agriculturae Boreali-Sinica
基金
财政部和农业农村部:国家现代农业产业技术体系资助(CARS-44-KXJ2)
山西省高校科技创新项目(2021L098)
山西省应用基础研究计划(青年)项目(20210302124360)
山西农业大学校青年科技创新项目(2019003)。
关键词
中华蜜蜂
SMOOTHENED
序列特征
生物信息学
表达特性
Apis cerana cerana
Smoothened
Sequence characterization
Bioinformatics analysis
Expression characteristics
