【摘要】 根据对中国野生葡萄华东葡萄株系白河-35-1在白粉病诱导下构建的cDNA文库中获得的1条EST序列设计特异引物,以总RNA逆转录产物为模板,进行SMART-RACE扩增。结合生物信息学方法对所获的序列进行开放阅读框、序列同源性分析,并预测了蛋白质的理化性质、信号肽、酶与非酶分析、亚细胞定位以及蛋白质二级结构。结果表明,获得的中国野生葡萄新基因的全长序列为854 bp,其开放阅读框为585 bp,5′非编码区为107 bp,3′非编码区为162 bp;同源性比对表明该新基因推导的氨基酸序列与拟南芥RNA结合蛋白和水稻推定的半乳糖基转移酶的同源性分别为55%和63%,第8~82位氨基酸序列与RNA识别基序相类似,是一个典型的结构功能域;推导该基因表达产物为相对分子质量为21 801.27、等电点为6.86的不稳定蛋白,定位于细胞核,不包含信号肽,二级结构主要是无规则卷曲。更多还原

【Abstract】 According to an EST sequence of cDNA libarary from Vitis pesudoreticulata Baihe-35-1 inoculated with Uncinula necator,a pair of specific primers were designed and the SMART-RACE were performed with reverse transcription products of total RNA as the template.Concurrently bioinformatic methods were applied to analyze the obtained sequence and its induced amino acid sequence,including ORF,homologous sequences,physical and chemical properties,signal peptide,enzyme/ non-enzyme,subcellular localization and protein secondary structure.The results showed that the full length cDNA had an open reading frame of 584 bp with 107 bp 5′ untranslated regions and 162 bp 3′ untranslated regions.The deduced amino acid had 55% identity to Arabidopsis RNA/DNA binding protein,63% identity to Oryza galactosyltransferase,and the 8 to 82 amino acid was homologous with RNA recognition motif,which belongs to a typical functional domain.The protein was instable,Mr=21 801.21,pI=6.86,without signal peptide,and located to nucleus.The secondary structure was primarily composed of randon coil.更多还原