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黑褐举腹蚁CrogOBP1基因克隆与表达分析

Cloning and Sequence Analysis of CrogOBP1 in Crematogaster rogenhoferi

  • 摘要: 本研究旨在克隆黑褐举腹蚁(Crematogaster rogenhoferi) CrogOBP1基因序列,通过生物信息学和分子生物学相结合的方法对其编码蛋白的信号肽、二级结构等进行预测分析。利用荧光定量RT-PCR检测CrogOBP1基因在黑褐举腹蚁头、触角、胸、腹和足中的表达相对丰度。结果显示,黑褐举腹蚁CrogOBP1基因开放阅读框全长为420 bp,编码139个氨基酸,具有20个氨基酸残基的信号肽。拥有6个保守的半胱氨酸位点等典型的气味结合蛋白的特点。预测编码蛋白质的分子量为15.4 kD,等电点为4.13。CrogOBP1与红胡须蚁(Pogonomyrmex barbatus)的OBP69a和扁胸切叶蚁属(Vollenhovia emeryi)的OBP69a聚为一支,亲缘关系较近。CrogOBP1基因在黑褐举腹蚁触角中表达量最高,且显著高于其它组织。本研究为进一步了解黑褐举腹蚁CrogOBP1基因功能和利用化学生态学手段防治橡副珠蜡蚧提供前期基础。

     

    Abstract: The aim of this study was to obtain the Cr ogOBP1 gene of Crematogaster rogenhoferi by cloning,through the method of bioinformatics and molecular biology to implement analytical prediction of the signal peptide and secondary structure. The expression specificity of CrogOBP1 in head, antennae, chest, abdomen and foot were determined by fluorescence quantitative RT-PCR. The results showed that the full-length of open reading frame of CrogOBP1 was 420 bp, encoding a signal peptide of 139 amino acids with 20 amino acid residues. It had six conserved cysteine sites and other characteristics of odorant binding proteins. It was predicted that the molecular weight of the encoded protein was 15.4 KD and the isoelectric point was 4.13. CrogOBP1 was closely related to OBP69 a of Pogonomyrmex barbatus and OBP69 a of Vollenhovia emeryi. It was highly expressed in the antennae which was significantly higher than other tissues. This study provide a theoretical basis for further understanding of the function and the control of Parasaissetia nigra Nietner by means of chemical ecology.

     

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