中华蜜蜂Malvolio基因Acmvl的克隆及组织表达分析  

Cloning and tissue expression profiling of Malvolio gene Acmvl in the Chinese honeybee,Apis cerana cerana(Hymenoptera: Apidae)

作　　者：孟娇[1] 马卫华 赵慧婷[3] 邵有全 田嵩浩 杨珊珊[1] 王树杰[1] 杜亚丽[1] 潘建芳[1] 姜玉锁[1] 

机构地区：[1]山西农业大学动物科技学院,山西太谷030801 [2]山西农业科学院园艺研究所,太原030031 [3]山西农业大学生命科学学院,山西太谷030801 [4]山西医科大学汾阳医学院,山西汾阳032200

出　　处：《昆虫学报》2015年第7期721-730,共10页Acta Entomologica Sinica

基　　金：国家公益性行业(农业)科研专项(201203080);国家自然科学基金项目(31272513)

摘　　要：【目的】本研究克隆了中华蜜蜂Apis cerana cerana Malvolio(Mvl)基因的c DNA序列,分析了其编码蛋白的结构特点,并探讨其mRNA在内勤蜂、采蜜蜂和采粉蜂各部位组织中的表达差异,以期为该基因的生物学功能研究提供参考。【方法】利用RT-PCR技术从中华蜜蜂内勤蜂头部组织中扩增和克隆获得Acmvl的全长序列,并采用多种生物信息学软件分析Acmvl蛋白的结构特征;采用Real-time PCR对中华蜜蜂Acmvl在内勤蜂、采蜜蜂和采粉蜂各组织中的表达特征进行分析。【结果】Acmvl基因c DNA全长为2 130 bp(Gen Bank登录号:KP662686),编码587个氨基酸,预测该蛋白分子量为65.86 k D,等电点为6.03,无信号肽,存在11个跨膜结构域、9个糖基化位点和14个潜在磷酸化位点;系统发育树分析结果显示,中华蜜蜂Acmvl与其他膜翅目昆虫Malvolio聚为一支,与小鼠Mus musculus和人Homo sapiens Nramp家族的Nramp2聚为另一大分支,且与小鼠、水稻Oryza sativa、黑腹果蝇Drosophila melanogaster和酵母Saccharomyces cerevisiae的Nramp家族同源体在跨膜区、跨膜区带电残基及转运蛋白特征结构域上有很高的保守性,尤其是与Nramp2。Acmvl基因在中华蜜蜂各部位组织中均有表达,但高表达于内勤蜂的胸部及采蜜蜂和采粉蜂的腹部和足部,提示该基因表达的差异影响采集行为。【结论】Acmvl属于Nramp基因家族,可能为Nramp2的同源基因,该基因影响采集行为可能与转运Cu2+,Mn2+和Fe2+(尤其是Fe2+)有关。[ Aim ] This study aims to clone the cDNA sequence of Malvolio （Mvl） gene in Apis cerana cerana, to analyze the structure of its coding protein, and to explore its expression profiles of mRNA in different tissues of nurse, nectar forager and pollen forager. This may provide a fundamental evidence for the future study of the physiological function of this gene. [ Methods ] The cDNA sequence of Acmvl was cloned from the brain of A. c. cerana nurse by reverse transcription polymerase chain reaction （RT- PCR）. A quantitative analysis of its expression level in different tissues （antenna, head, thorax, abdomen and leg） of the nurse, nectar forager and pollen forager was conducted using real-time PCR.[ Results] The full-length cDNA sequence of Acmvl of A. c. cerana is 2 130 bp （GenBank accession no. KP662686）, encoding 578 amino acids with the predicted MW and pI of 65. 86 kD and 6. 03, respectively. The encoded protein has no signal peptide but has 11 transmembrane domains, 9 predicted glycosylation sites and ld potential phosphorylation sites. Phylogenetic analysis indicated that Acmvl of A. c. cerana and Malvolio proteins of other Hymenoptera insects gather in the same clade, and Acmvl of A. c. cerana and Nramp2 proteins of Nramp family from Mus musculus and Homo sapiens gather in another clade. Acmvl of A. c. cerana has an extensive similarity to Nramp proteins of Mus musculus, Oryza sativa, Drosophila melanogaster and Saccharomyces cerevisiae in transmembrane domain （TM）, charged residues of TM and transport motif, especially with Nramp2 proteins. The Acmvl transcript was clearly detected in different tissues of the worker, and was expressed significantly higher in thorax of nurse, abdomen and legs of nectar and pollen forager, suggesting that the expression of Acmvl gene has an effect on foraging behavior of bees. [ Conclusion ] As a member of the family of Nramp, Acmvl gene may be homologous with Nramp2, and it affects foraging behavior in part because of its involvement in transportation of

关 键 词：中华蜜蜂 Malvolio 基因克隆 序列分析 组织表达模式 

分 类 号：Q966[生物学—昆虫学]