橘小实蝇半胱氨酸蛋白酶基因克隆及表达模式

杨文佳; 许抗抗; 王进军; 李灿

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橘小实蝇半胱氨酸蛋白酶基因克隆及表达模式

研究论文 | 更新时间：2023-12-11

- 橘小实蝇半胱氨酸蛋白酶基因克隆及表达模式
- Molecular cloning and expression profiling of a cysteine proteinase gene in the oriental fruit fly Bactrocera dorsalis (Hendel)
- 中山大学学报(自然科学版)(中英文) 2017年56卷第1期页码：131-137
- 作者机构：
  
  1. 贵阳学院生物与环境工程学院//有害生物控制与资源利用贵州省高校特色重点实验室,贵州,贵阳,550005
  2. 
- 作者简介：
- 基金信息：
- DOI：
  中图分类号：
- 纸质出版日期：2017，
  
  网络出版日期：2017-1-25，
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杨文佳, 许抗抗, 王进军, 等. 橘小实蝇半胱氨酸蛋白酶基因克隆及表达模式[J]. 中山大学学报(自然科学版)(中英文), 2017,56(1):131-137.

YANG Wenjia, XU Kangkang, WANG Jinjun, et al. Molecular cloning and expression profiling of a cysteine proteinase gene in the oriental fruit fly Bactrocera dorsalis (Hendel)[J]. Acta Scientiarum Naturalium Universitatis SunYatseni, 2017,56(1):131-137.
杨文佳, 许抗抗, 王进军, 等. 橘小实蝇半胱氨酸蛋白酶基因克隆及表达模式[J]. 中山大学学报(自然科学版)(中英文), 2017,56(1):131-137. DOI：

YANG Wenjia, XU Kangkang, WANG Jinjun, et al. Molecular cloning and expression profiling of a cysteine proteinase gene in the oriental fruit fly Bactrocera dorsalis (Hendel)[J]. Acta Scientiarum Naturalium Universitatis SunYatseni, 2017,56(1):131-137. DOI：

摘要

为了深入了解半胱氨酸蛋白酶基因的作用和寻求害虫防治新的分子靶标，利用RT-PCR和RACE技术，从橘小实蝇Bactrocera dorsalis (Hendel)体内克隆获得半胱氨酸蛋白酶基因Bdcyp的cDNA全长序列（GenBank登录号：KU904503），采用生物信息学软件分析了Bdcyp编码蛋白的特性，利用实时定量PCR技术检测其在不同发育阶段和不同组织的表达量。橘小实蝇Bdcyp的cDNA全长为1 955 bp，开放阅读框为1 653 bp，编码551个氨基酸，预测蛋白质相对分子质量和等电点分别为62 900和6.11。Bdcyp具有半胱氨酸蛋白酶的典型特征，即信号肽、3个催化活性区域（Cys-His-Asn）以及2个保守的ERFNIN和GNFD特征基序。系统发育分析表明Bdcyp编码的蛋白质与橄榄实蝇半胱氨酸蛋白酶亲缘关系最近。Bdcyp在三龄幼虫、蛹和成虫期均有表达，其在蛹中期的表达量最高；表达部位主要在幼虫脂肪体，其次为气管和表皮，而在中肠的表达量最低。这些结果为进一步研究Bdcyp基因功能提供了依据。

Abstract

In order to better understand the roles of cysteine proteinase gene and develop a novel molecular target for insect pest control

we obtained the full-length cDNA sequence of Bdcyp (GenBank accession no. KU904503) from the oriental fruit fly

Bactrocera dorsalis (Hende) by using RT-PCR and RACE methods. The deduced amino acid sequence analysis of Bdcyp was performed by using bioinformatics methods. The expression patterns of Bdcyp in different stages and tissues were investigated using quantitative real-time PCR (qPCR). The Bdcyp was 1 955 bp in lenght

containing an open reading frame of 1 653 bp which encodes 551 amino acid residues with a predicated molecular weight of 62 900 and isoelectric point of 6.11. Bdcyp had typical features of cysteine proteinase

including signal peptide

three catalytic active regions (Cys-His-Asn)

and two conserved motifs (ERFNIN and GNFD). Phylogenetic analysis showed that Bdcyp has the closest genetic relationship with the cysteine proteinase of B. oleae. Bdcyp was constitutively expressed in the tested stages

including third-instar larvae

pupae and adults. The highest expression level was observed at the middle of pupal stage. Tissuespecific expression showed that the highest expression was in the fat body

followed by trachea and integument

and the expression in midgut was the lowest. The results provide the basis for further studying the function of Bdcyp.

关键词

橘小实蝇半胱氨酸蛋白酶克隆序列分析表达模式

Keywords

Bactrocera dorsaliscysteine proteinasecloningsequence analysisexpression pattern

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