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 vitellogenincDNA in the bumblebee，Bombus ignitus
李继莲1**,吴杰1,彩万志2,彭文君1***
点击：2096次 下载：6次
DOI：
作者单位：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
中文关键词：熊蜂，Bombus ignitus，卵黄原蛋白，荧光定量RT-PCR
英文关键词：bumblebee，Bombus ignitus，vitellogenin，quantitative real-time RT-PCR
中文摘要： 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。
英文摘要： 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 acidswithin a putative signal peptide of 16 residues (GenBank accession number FJ913883) ． Close to the C-terminus there is aGL / 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 ofconserved motifs． The mRNA of B． ignitus vitellogenin(Vg)was analysed in queens，workers and drones，from pupae toadults． Vg mRNA was found in the white-eyed pupal ( Pw) stage in queens and its expression increased during the entirepupal 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 andreaching a high point in 15 d，after which it decreased progressively． Vg mRNA was also observed in drones，the highestlevel of expression being observed in newly emerged adult drones．