›› 2013, Vol. 56 ›› Issue (6): 584-593.

• 研究论文 • 上一篇    下一篇

烟粉虱MEAM1隐种卵黄原蛋白受体基因cDNA的克隆、 序列分析及在不同发育时期的表达

程璐, 郭建洋, 刘树生, 叶恭银*   

  1. (浙江大学昆虫科学研究所, 农业部农业昆虫学重点实验室, 杭州 310058)
  • 出版日期:2013-06-20 发布日期:2013-06-20

Molecular cloning, sequence analysis and developmental expression profile of vitellogenin receptor gene in the whitefly Bemisia tabaci Middle East-Asia Minor 1 (Hemiptera: Aleyrodidae)

CHENG Lu, GUO Jian-Yang, LIU Shu-Sheng, YE Gong-Yin*   

  1. (Key laboratory of Agricultural Entomology, Ministry of Agriculture, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, China)
  • Online:2013-06-20 Published:2013-06-20

摘要: 卵黄原蛋白受体(vitellogenin receptor, VgR)是卵黄原蛋白被卵母细胞摄取的关键因子, 在卵黄发生和卵母细胞发育等生理过程中发挥着重要作用。为探讨烟粉虱Bemisia tabaci VgR的功能, 我们采用RT-PCR和RACE等技术扩增了烟粉虱MEAM1隐种B. tabaci Middle East-Asia Minor 1 (MAEM1) 的VgR基因cDNA 全长序列。生物信息学分析表明, 烟粉虱MEAM1隐种的VgR基因cDNA全长5 774 bp, 编码1 919个氨基酸, 推测分子量约201 kDa, N-端前31个氨基酸为信号肽。烟粉虱MEAM1隐种的VgR属于低密度脂蛋白受体(low density lipoprotein receptor, LDLR)家族, 蛋白质三维结构预测分析表明, 该受体具有LDLR家族基因典型的保守功能结构域。通过实时荧光定量PCR技术研究了烟粉虱MEAM1隐种VgR基因不同发育时期的表达, 结果表明VgR基因在伪蛹期开始表达, 并在羽化后1 d达到高峰, 此后逐渐降低, 3 d后又逐渐升高, 直至羽化后7 d达到峰值。研究结果丰富了卵黄原蛋白受体家族基因的数据库, 为今后深入研究并揭示烟粉虱卵黄发生的调控机制奠定了基础。

关键词: 烟粉虱, 卵黄原蛋白受体, 基因克隆, 序列分析, 表达谱

Abstract: Vitellogenin receptor (VgR) is one of the key factors during the uptake of vitellogenin (Vg) by oocytes, and plays a critical role in vitellogenesis and oocyte development. In order to define the physiological function of VgR, VgR cDNA in the whitefly, Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) was sequenced using the combined methods of reverse transcription PCR (RT-PCR) and rapid amplification of cDNA ends (RACE). The bioinformatical analysis demonstrated that the full-length cDNA of VgR from B. tabaci MEAM1 is 5 774 bp in size. The putative mature VgR has 1 919 amino acids with a molecular weight of 201 kDa. The signal peptide at the N-terminal end contains 31 amino acids. The VgR of this whitefly has all the typical conserved domains of the low density lipoprotein receptor (LDLR) family. The expression profile of VgR gene at different developmental stages of the whitefly was detected by realtime quantitative PCR. The results showed that the expression of VgR gene was initiated at the pseudo-pupal period and increased rapidly on the 1st day after eclosion and reached the peak level on the 7th day after eclosion. These results will enrich the gene database of VgR and provide valuable information to ascertain the regulation mechanism of vitellogenesis in B. tabaci.

Key words: Bemisia tabaci, vitellogenin receptor, gene cloning, sequence analysis, expression profile