红光熊蜂卵黄原蛋白基因的cDNA全长序列克隆和表达分析  

The expression of the mRNA and cloning for the vitellogenin cDNA in the bumblebee,Bombus ignitus

作　　者：李继莲[1] 吴杰[1] 彩万志[2] 彭文君[1] 

机构地区：[1]农业部授粉昆虫生物学重点实验室中国农业科学院蜜蜂研究所,北京100093 [2]中国农业大学昆虫系,北京100193

出　　处：《应用昆虫学报》2012年第5期1132-1139,共8页Chinese Journal of Applied Entomology

基　　金：国家自然基金(30972149);国际科技合作项目(2009DFA32600);公益性行业(农业)科研专项课题(201203080-4);国家蜂产业技术体系建设专项(CARS-45)

摘　　要：红光熊蜂Bombus ignitus Smith是许多经济作物和野生植物的重要授粉昆虫之一。卵黄原蛋白基因(vitellogenin,Vg)在昆虫的生殖调控中和行为方面起到重要的作用,本试验对Vg基因全长cDNA的克隆和测序及在蜂王、工蜂和雄性蜂三型蜂中的表达分析得出:Vg基因的全长cDNA为5 481 bp,GenBank中的登录号为FJ913883,有一个完整的开放阅读框(ORF),编码1 772个氨基酸,N-末端的前16个氨基酸为一个信号肽。接近C-末端区域存在保守的GL/ICG基元,其后含有9个半胱氨酸,而且DGXR位于GL/ICG基元上游18个氨基酸残基处。其氨基酸序列与韩国的熊蜂B.ignitus和B.hypocrita相似性高达95%,与西方蜜蜂Apis mellifera的相似性达到51%。Vg的mRNA首先在蜂王蛹期的白眼蛹(Pw)时期出现,其表达量在蜂王整个蛹期发育过程中呈上升趋势,且在黑眼蛹(Pbd)时期达到最高,在成年蜂的脂肪体中的表达量仍在升高,甚至更高。Vg也在工蜂蛹期的白眼蛹(Pw)时期被检测到,然后在整个蛹期发育过程中呈现上升趋势,在刚羽化出房时达到高峰,Vg的mRNA水平随着成年蜂日龄的增加而增加,到15日龄时达到最高,然后呈现下降趋势。对于雄性蜂,Vg的mRNA虽然卵黄原蛋白基因的mRNA水平几乎在整个蛹期发育阶段都表达,但是表达水平非常低,只有在刚羽化出房时期表达水平较高。Bombus ignitus Smith is one of the important pollinators of many economically important crops and wildflowers. vitellogenin （Vg） plays a key role in the control of reproduction and reproductive behavior of this species. In this study, Vg cDNA of B. ignitus was cloned and sequenced. It is composed of 5 481 bp and contains an ORF of 1 772 amino acids within a putative signal peptide of 16 residues （GenBank accession number FJ913883）. Close to the C-terminus there is a GL/ICG motif followed by nine cysteines, and a DGXR motif is located 18 residues upstream of the GL/ICG motif. BLASTX analysis of the conceptual translation products revealed 95% similarity to B. ignitus from Korea and to B. hypocrita. The deduced amino acid sequence shows significant similarity with Apis mellifera （51% ） and a high number of conserved motifs. The mRNA of B. ignitus vitellogenin（Vg）was analysed in queens, workers and drones, from pupae to adults. Vg mRNA was found in the white-eyed pupal （Pw） stage in queens and its expression increased during the entire pupal development, reaching a peak in the dark brown pupal （Pbd） stage. It also had a high expression in the adult fatbody. In workers, the Vg expression was detected in the white-eyed pupal （Pw）stage, its levels increasing with age and reaching a high point in 15 d, after which it decreased progressively. Vg mRNA was also observed in drones, the highest level of expression being observed in newly emerged adult drones.

关 键 词：熊蜂 BOMBUS ignitus 卵黄原蛋白 荧光定量RT-PCR 

分 类 号：S891[农业科学—特种经济动物饲养] Q963[农业科学—畜牧兽医]