›› 2015, Vol. 58 ›› Issue (4): 361-366.

• 研究论文 •    下一篇

东亚飞蝗UBX结构域包含蛋白基因 LmUBX2 的克隆和表达分析

冉永红, 黄婷, 郝友进, 陈斌, 何正波*   

  1. (重庆师范大学昆虫与分子生物学研究所, 重庆 400047)
  • 出版日期:2015-04-20 发布日期:2015-04-20
  • 作者简介:冉永红, 女, 1990年生, 重庆万州人, 硕士研究生, 研究方向为昆虫分子生物学, E-mail: 1048765288@qq.com

Cloning and expression analysis of a UBX domain-containing protein gene LmUBX2 from the migratory locust, Locusta migratoria manilensis (Orthoptera: Locustidae)

RAN Yong-Hong, HUANG Ting, HAO You-Jin, CHEN Bin, HE Zheng-Bo*   

  1. (Institute of Entomology and Molecular Biology, Chongqing Normal University, Chongqing 400047, China)
  • Online:2015-04-20 Published:2015-04-20

摘要: 【目的】UBX结构域包含蛋白是p97/CDC48的辅助因子。p97在泛素化相关的多种细胞过程中起着重要的作用,如依赖泛素蛋白酶体系统的蛋白质降解和同型膜融合等。本研究旨在克隆东亚飞蝗 Locusta migratoria manilensis (Meyen)的UBX结构域包含蛋白基因,分析其组织和发育表达格局,为进一步研究UBX结构域包含蛋白基因的功能奠定基础。【方法】通过分析东亚飞蝗的转录组数据克隆UBX结构域包含蛋白基因,采用实时定量PCR技术分析该基因在不同发育时期和成虫不同组织中的表达水平。【结果】克隆到东亚飞蝗的一个UBX结构域包含蛋白基因,命名为 LmUBX2LmUBX2 开放阅读框长1 020 bp,编码399个氨基酸,预测分子量和等电点分别为37.8 kDa和6.03,与其他UBX结构域包含蛋白的氨基酸一致性为37%~64%,N端和C端分别有一个保守的UBA结构域和UBX结构域。序列比较和系统发育分析发现 LmUBX2 属于SAKS1亚家族。定量分析发现,LmUBX2 在整个生命周期中都有表达,但成虫期的表达水平最高;在检测的所有组织中都有表达,但在精巢和卵巢中表达水平最高。【结论】研究结果说明 LmUBX2 可能参与东亚飞蝗多种生理过程,尤其可能与东亚飞蝗的生殖有关,但还需深入研究。

关键词: 东亚飞蝗, UBX结构域包含蛋白, LmUBX2, 基因克隆, 表达模式

Abstract: 【Aim】 Ubiquitin regulatory X (UBX) domain-containing proteins function as cofactors of p97/CDC48 which is involved in multiple ubiquitin-related processes including protein degradation through the ubiquitin-proteasome system and homotypic membrane fusion. Our study aims to clone the UBX domain-containing protein gene from Locusta migratoria manilensis, and to analyze its expression patterns in different tissues and at different developmental stages, so as to provide the foundation for further function research. 【Methods】 A transcriptome database for L. migratoria manilensis was mined through bioinformatic analysis. Gene expression levels were analyzed in different tissues of L. migratoria manilensis adults and at different developmental stages using real-time PCR. 【Results】 A UBX domain-containing protein gene was identified, and named LmUBX2. The open reading frame (ORF) encodes a protein of 339 amino acids with a predicted molecular weight of 37.8 kDa and a theoretical pI of 6.03. BLAST analysis showed that it shares 37%-64% amino acid sequence identities to other UBX domain-containing proteins. LmUBX2 possesses a conserved N-terminal UBA domain and a C-terminal UBX domain. Sequence comparison and phylogenetic analysis revealed that LmUBX2 is a member of the SAKS1 subfamily. Gene expression analysis results showed that LmUBX2 was expressed during the whole life cycle, and highly expressed in ovary and testis. 【Conclusion】 These findings suggest that LmUBX2  might be involved in multiple physiological processes of L. migratoria manilensis. Especially, LmUBX2 might be related to the reproduction of L. migratoria manilensis, although further research needs to be performed to confirm this correlation.

Key words: Locusta migratoria manilensis, UBX domain-containing protein; LmUBX2, gene cloning, expression pattern