| 摘要: |
| 【目的】在受白粉病菌侵染的中国野葡萄叶片中克隆锌指蛋白基因全长，研究葡萄抗白粉病的分子机制。【方法】在前期采用mRNA差异显示技术获得的中国野葡萄华东葡萄株系白河35-1抗白粉病基因表达差异cDNA片段T11 GG/B0324-292的基础上，利用cDNA末端快速扩增（RACE）技术克隆了该cDNA全长序列，并结合生物信息学方法对所获取的序列进行分析。【结果】该cDNA全长2 663 bp(GenBank登录号HM008621)，具有完整的开放阅读框，基因全长2 223 bp，编码1个由740个氨基酸组成的锌指蛋白（Zinc finger protein），5′和3′非翻译区长度分别为395和44 bp。多重比较分析表明，该基因推导的氨基酸序列与拟南芥、蒺藜苜蓿、水稻的C3H亚型CCCH型锌指蛋白的同源性分别为61%，70%和60%。【结论】利用RACE技术获得了锌指蛋白基因全长，为进一步研究锌指蛋白基因与葡萄抗白粉病的关系奠定了基础。 |
| 关键词: 中国野葡萄 白粉病 锌指蛋白 转录因子 RACE 生物信息学 |
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| 基金项目:国家自然科学基金项目(30871701,30671446)；教育部新世纪优秀人才支持计划项目 |
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| Cloning and bioinformatic analysis of zinc finger protein gene in Chinese wild Vitis pseudoreticulata |
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| Abstract: |
| 【Objective】 A full length cDNA of a zinc finger protein gene was cloned in Chinese wild Vitis pseudoreticulata which were infected by Uncinula necator(Schw.)Burr,and the molecular mechanisms of grape resistant to powdery mildew were studieds.【Method】This article was based on T11 GG/B0324-292 cDNA fragment of Chinese wild Vitis pseudoreticulata Baihe 35-1,the cDNA fragment came from mRNA differential display technology,the full-length cDNA of the fragment was cloned by rapid amplification of cDNA ends(RACE) technique.Concurrently bioinformatic methods were applied to analyze the obtained sequence.【Result】The cDNA sequence consisted of 2 663 bp(GenBank accession number HM008621)and had an open reading frame (ORF) of 2 223 bp,encoded a zinc finger protein of 740 amino acids,the non-translation of 5′and 3′-terminal region was 395,44 bp respectively.The multiple comparison sequence analysis showed that the deduced amion acid sequences shared identity to 61%,70%and 60%with A.sthaliana,M.truncatula and O.sativa,respectively.【Conclusion】The full-length zinc finger protein gene has been cloned by RACE technique,which lays the foundation for further research on the relationship between the gene and grape resistantance to powdery mildew. |
| Key words: Chinese wild grape powdery mildew zinc finger protein transcription factor RACE bioinformatics |
