【作者】 包丽媛；

【导师】 侯和胜；

【作者基本信息】 辽宁师范大学 ， 植物学， 2009， 硕士

【摘要】 根据NCBI数据库相关信息,我们设计了特异引物,从野生夏葡萄叶片克隆了PR3和PR5基因。利用公共数据库数据,采用生物信息学的理论和方法,对获得的野生夏葡萄病程相关蛋白基因进行了较全面的性质分析。为后续的功能研究提供理论基础,也为利用生物信息学方法深入研究功能基因提供新的思路。本文主要研究结果如下:1.根据数据库信息,成功克隆出野生夏葡萄PR3和PR5基因。2.采用生物信息学方法进行信号肽分析,结果表明,PR3很可能是具有信号肽的分泌型蛋白,而PR5具有信号肽的可能性很小。3.运用生物信息学的方法和工具对野生夏葡萄PR3和PR5基因的核苷酸和氨基酸序列进行分析;并对它们的疏水性/亲水性、跨膜结构域、蛋白质二级结构、三级结构和功能结构域进行预测和分析。这些预测和分析结果表明:野生夏葡萄PR3的功能域是chitinase,PR5的功能域是thaumatin。4.NCBI上的电子表达谱分析表明野生夏葡萄PR3和PR5可能在葡萄多种器官中表达,但不同的PR基因在不同的组织中表达强弱不同。5.用PlantCARE分析野生夏葡萄PR3和PR5基因的启动子,发现它们具有启动子的基本元件TATA-box、CAAT-box,并含有真菌诱导元件Box-W1及与防御和胁迫相关的作用元件TC-rich repeats,还包含与水杨酸相关的作用元件TGA-element,与脱落酸相关的作用元件ABRE,与茉莉酸甲酯相关的作用元件CGTCA-motif和TGACG-motif等很多顺式作用元件。这些顺式作用元件的存在,表明PR3和PR5基因的表达可能受到不良环境和植物激素的诱导和调节。更多还原

【Abstract】 According to the data in NCBI,we designed primers and cloned PR3 and PR5 genes from Vitis aestivalis.Based on the public databases,we conducted comprehensive analysis of PR3 and PR5 by using the methods of bioinformatics.It will provide some useful information for its following function research and new methods for the researches of other gene families.The results of this study are as follows:1.According to the information from NCBI,we successfully cloned PR3 and PR5 genes from V.aestivalis.2.The results of signal peptide analysis indicated that PR3 has signal peptide sequence while PR5 may have no signal peptide sequence.3.By the bioinformatics methods and tools,the composition and sequences of amino acids and nucleotides,hydrophobicity and hydrophilicity, transmembrane domains,the secondary and tertiary structures of PR3 and PR5 were analyzed and predicted. The results showed that PR3 contains chitinase functional domain and that PR5 contains thaumatin functional domain.4.The predicted expression profiles showed that these two genes might be expressed in most grape organs.However, the different PR was expressed in different organs at quite different levels.5.The functional cis-acting elements in the promoters of PR3 and PR5 gene were analyzed by using PlantCARE software.Many cis-acting elements were found such as: TATA-box;CAAT-box;Box-W1,a fungal elicitor responsive element;TC-rich repeats,a defense and stress responsive element;TGA element,an auxin-responsive element;ABRE,an abscisic acid responsive element;CGTCA-motif and TGACG-motif, MeJA-responsive elements and so on.更多还原