›› 2010, Vol. 53 ›› Issue (5): 487-496.doi:

• 研究论文 • 上一篇    下一篇

苜蓿盲蝽气味结合蛋白基因Alin-OBP1的克隆及表达谱分析

谷少华,张雪莹,张永军,吴孔明,郭予元   

  • 出版日期:2010-05-20 发布日期:2010-05-20
  • 通讯作者: 张永军

Cloning and expression pattern analysis of an odorant binding protein gene Alin-OBP1 in the lucerne plant bug, Adelphocoris lineolatus (Goeze) (Hemiptera: Miridae)

GU Shao-Hua, ZHANG Xue-Ying, ZHANG Yong-Jun, WU Kong-Ming, GUO Yu-Yuan   

  • Online:2010-05-20 Published:2010-05-20

摘要: 有证据表明昆虫气味结合蛋白(odorant binding proteins, OBPs)与其嗅觉识别密切相关,起着运输外界脂溶性气味分子通过嗅觉感器淋巴液到达嗅觉受体的关键作用.为了更好地了解OBPs在苜蓿盲蝽Adelphocoris lineolatus (Goeze)嗅觉识别中的作用,本研究首次克隆了苜蓿盲蝽气味结合蛋白基因Alin-OBP1 (GenBank序列号GQ477022). 测序和序列分析结果表明,该基因开放阅读框长438 bp, 编码145个氨基酸,预测分子量为15.69 kDa,等电点为5.01,N-末端疏水区包含由18个氨基酸组成的信号肽.蛋白特征分析表明,该基因翻译后的蛋白质具有昆虫气味结合蛋白的典型特征,即氨基酸序列中有6个保守的半胱氨酸残基.利用RT-PCR和Real-time PCR技术对Alin OBP1在苜蓿盲蝽成虫不同组织和各个发育阶段的表达水平进行了测定,结果显示Alin-OBP1几乎全部在触角中表达.不同发育阶段Alin-OBP1表达量不同,在5龄若虫和成虫阶段表达水平最高.结果提示Alin-OBP1可能在苜蓿盲蝽感受包括性信息素在内的外界化合物的过程中发挥着重要作用.

关键词: 苜蓿盲蝽, 气味结合蛋白, 基因克隆, 表达谱分析, 发育阶段

Abstract: In insects,there is now increasing evidence that the odorant binding proteins ( OBPs) are
involved in olfaction perception and play a key role in transporting hydrophobic odorants across the sensillum
lymph to the olfactory receptors ( ORs) . In order to better understand the role of OBPs in the olfactory
system of the lucerne plant bug,Adelphocoris lineolatus ( Goeze) ,we cloned a cDNA encoding odorant
binding protein 1 of A. lineolatus in the laboratory,which was named Alin-OBP1 ( GenBank accession no.
GQ477022) . The results of sequencing and sequence analysis showed that the open reading frame ( ORF)
of Alin-OBP1 was 438 bp in length,encoding 145 amino acids,the predicted molecular weight ( MW) was
15. 69 kDa,and the isoelectric point ( pI ) was 5. 01. The predicted N-terminal hydrophobic region
containing 18 amino residues displayed the characteristic features of a signal peptide. Protein signature
analyses revealed that it shared typical structural features of odorant binding proteins with other insects,
including six conservative cysteine residues. By RT-PCR and Real-time PCR methods,we determined the
expression pattern of Alin-OBP1 in different adult tissues and developmental stages of A. lineolatus. It was
found that Alin-OBP1 was dominantly expressed in the antenna of A. lineolatus. In addition,Alin-OBP1
was expressed in various nymphal and adult stages at different expression levels and had higher expression
level in the 5th instar nymphal and adult stages. The results suggest that Alin-OBP1 may play a key role in
the perception of sex pheromones and other chemical odorants in A. lineolatus.

Key words: Adelphocoris lineolatus, odorant binding protein, gene cloning, expression pattern analysis, developmental stage