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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (19): 1-8.doi: 10.11924/j.issn.1000-6850.2013-0368

所属专题： 生物技术； 园艺

• 林学 园艺 园林 •    下一篇

巴西橡胶树HbUBC22基因克隆与表达分析

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  • 收稿日期:2013-02-01 修回日期:2013-03-18 出版日期:2013-07-05 发布日期:2013-07-05
  • 基金资助:
    中国热带农业科学院橡胶研究所人才项目

Cloning and expression of HbUBC22 in Hevea brasiliensis Muell. Arg.

  • Received:2013-02-01 Revised:2013-03-18 Online:2013-07-05 Published:2013-07-05

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摘要： 泛素蛋白酶体途径（ubiquitin proteasome pathway, UPP）是细胞内最重要的且具有高度选择性的蛋白质降解途径，而泛素结合酶E2/UBC是该途径的一个关键酶。本研究通过对巴西橡胶树同源EST序列拼接和RT-PCR扩增的方法，克隆了一个巴西橡胶树的泛素结合酶基因HbUBC22，该基因的开放阅读框（ORF）共807 bp，编码268个氨基酸，编码蛋白含有一个保守的UBC结构域和一个高度保守的半胱氨酸催化位点。系统进化关系分析发现，HbUBC22蛋白与蓖麻、杨树、油茶和葡萄的UBC蛋白的同源性最高，分别为89%、85%、85%和81%；在拟南芥基因组中，HbUBC22则与AtUBC22的同源性最高达到73%。表达谱分析结果显示，HbUBC22受乙烯利和茉莉酸甲酯诱导显著上调表达。原核表达分析结果表明，经IPTG诱导后，HbUBC22能够在大肠杆菌中表达出目标蛋白。本研究结果为进一步研究HbUBC22在橡胶树中的生物学功能奠定了良好基础。

关键词: 表达, 表达

Abstract: The ubiquitin proteasome pathway (UPP) is the crucial regulatory mechanism for selective protein degradation. Ubquitin-conjugating enzyme (UBC/E2) is a key enzyme involved in UPP. In this study, the HbUBC22 was cloned in Hevea brasiliensis by assembly the Hevea homologous EST and RT-PCR amplification. The ORF of HbUBC22 contained 807 bp in length and encoded 268 amino acid residues. The putative protein of HbUBC22 had a conserved UBC domain and a conserved active-site cysteine in the UBC domain. Phylogenetic analysis showed that HbUBC22 shared the highest similarity with the UBC protein from Ricinus communis, Populus trichocarpa, Camellia oleifera, and Vitis vinifera, with 89%, 85%, 85% and 81% sequence identities, respectively. In the genome of Arabidopsis thaliana, HbUBC22 shared the highest similarity with AtUBC22 with 73% identical residues. Transcription pattern analysis revealed that HbUBC22 expression was markedly induced by ethephon (ET) and methyl jasmonate (MeJA). Prokaryotic expression in E. coli indicated that HbUBC22 recombinant protein was expressed in the IPTG induced cells. These results provide a good basis for further characterization of the biological function of HbUBC22 in Hevea brasiliensis.

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