摘要黄瓜(Cucumis sativus L.)不仅作为我国农村栽培面积最大的蔬菜作物，而且也是世界上种植广泛的经济作物，它为农民带来较高的经济效益从而带动了农村的建设发展，因此关于黄瓜的相关调控理论的研究一直受到人们的重视。黄瓜全基因组序列测定已经完成这就为后续的基因组学研究奠定了基础。本论文就是以从黄瓜基因序列出发，采用生物信息学手段对黄瓜中与拟南芥生长素受体基因序列具有很高同源性的基因序列展开研究。我们之所以预测这2个基因可能就是一直寻找无果的黄瓜中的生长素受体基因，并将它们暂命名为 CsTIR 和CsAFB，是因为这两条基因序列与拟南芥中生长素受体基因序列有较高的相似性。为研究这2个基因的功能，我们提取黄瓜的RNA，并以其为模板通过逆转录得到cDNA，设计特异性PCR引物从而来克隆这2个基因用于后续研究和分析。我们对CsTIR 和 CsAFB 编码的蛋白质的功能域进行分析，以及运用软件分析了进化保守性，并且利用半定量RT-PCR对CsTIR和CsAFB基因在不同品种、不同组织及不同时期的黄瓜中的表达模式进行了研究，检测其相对表达丰度。本论文的研究成果将为证实黄瓜生长素受体基因就是CsTIR和CsAFB提供最直接的证据，并且这也是将来研究CsTIR和CsAFB基因的其信号通路调控机制及其在黄瓜中的具体的生物学功能提供理论依据。47891

毕业论文关键词：黄瓜；生长素；基因克隆；表达模式

Abstract

Cucumber (Cucumis sativus L.) is one of the important horticultural and economic crops in the world，which is the first large area of protected cultivation of vegetable crops in China and contributing substantially to rural economic development，thus，the research on the basic theory of cucumber has been widely appreciated. As the whole genome sequences of cucumber had been published, it laid the foundation for the subsequent genome research. This work is based on the genome sequences of cucumber. Compared with the sequences of auxin receptor genes in Arabidopsis, two highly conserved sequences were selected. We predict that these two genes might be the auxin receptor genes in cucumber, and named them of CsTIR and CsAFB. In order to study the function of these genes, we extracted the RNA of cucumber and obtained cDNA by reverse transcription, used specificly designed PCR primers to clone this 2 genes for follow-up analysis. The functional protein domains of CsTIR and CsAFB were predicted by bioinformatics methods. Phylogenetic and conservation analysis were taken with sequences from different plants. Expression patterns of the two genes in different varieties, different tissues and growing period were analyzed by semi-q RT-PCR. This study of CsTIR and CsAFB provided a strong basis of cucumber auxin receptor research, which is also contributed to the  research of their function and regulatory mechanism in the future.

Keyword: Cucumber；Auxin；Gene cloning；Expression pattern

 目    录

1 前    言 4

2 黄瓜生长素受体同源基因寻找 4

2.1 黄瓜基因组的序列对比 4

2.2 CsTIR和CsAFB基因的进化分析 7

3 CsTIR和CsAFB基因在黄瓜中的表达模式分析 8

3.1 实验材料与试剂 8

3.2 实验方法 8

3.3 实验结果 11

3.4 讨论 12

4 总结与展望 12

参考文献 14

致  谢