目录
摘要Ⅱ
关键词Ⅱ
AbstractⅢ
引言
引言1
1□材料1
2□方法2
2.1□克隆目的基因2
2.1.1□引物设计2
2.1.2□PCR克隆目的基因2
2.1.3□目的基因的检测2
2.1.4□PCR产物胶回收3
2.1.5□配制LB和1/2 MS培养基3
2.1.6□连接中间载体4
2.1.7□转化大肠杆菌DH5α感受态细胞4
2.1.8□生物信息学分析4
2.2□重组表达载体构建5
2.2.1□设计接头引物5
2.2.2□连接接头引物与目的基因5
2.2.3□目的基因连接表达载体并转化大肠杆菌5
2.2.4□菌液PCR检测5
2.2.5□大肠杆菌质粒提取6
2.2.6□转化农杆菌GV31017
2.3□农杆菌侵染转化拟南芥7
2.4□T0代种子筛选并继繁8
3□结果与分析8
3.1□目的基因的克隆8
3.2□过表达载体的构建9
3.3□大肠杆菌菌液PCR检测9
3.4□农杆菌菌液PCR检测10
3.5□拟南芥生长状况分析10
3.6□FTa1和FTa2蛋白的功能初探11
4□讨论12
致谢14
参考文献14
蚕豆(Vicia faba L.)成花素基因FTa1和FTa2
的同源克隆与分析
摘 要
为探究FLOWERING LOCUS T(FT)基因在植物开花过程中的作用，深入研究蚕豆开花的分子机理，本实验对蚕豆成花素基因FTa1和FTa2基因进行了克隆和分析。根据与蚕豆同源性较高的豌豆FTa1和FTa2基因序列设计引物，以蚕豆叶片cDNA为模板，扩增得到FTa1和FTa2基因。运用重组法构建植物表达载体，通过农杆菌转化法侵染开花时期的野生型和ft10突变体拟南芥，鉴定蚕豆FT基因在植物开花中的功能。结 *51今日免费论文网|www.jxszl.com +Q: @351916072@ 
果表明，FTa1基因全长为216bp，FTa2基因全长为531bp，FTa2基因的过表达能够显著促进拟南芥开花，而FTa1基因不能促进开花，这可能是由于FTa1蛋白是一个失去功能的截断蛋白，暗示了蚕豆进化中重复基因的去功能化。
HOMOLOGOUS CLONING AND ANALYSIS OF THE FLORIGEN GENES FTa1 AND FTa2 FROM FABABEAN
ABSTRACT
In order to explore the role of FLOWERINGLOCUST (FT) gene in plant flowering and further study the molecular mechanism of fababean flowering, FTa1 and FTa2 genes were cloned and analyzed. According to the sequence of pea FTa1 and FTa2 genes with high homology to fababean, primers were designed and FTa1 and FTa2 genes were amplified from cDNA of fababean leaves. The plant expression vector was constructed by recombination method, and the wild type and ft10 mutant Arabidopsis thaliana were infected by Agrobacterium tumefaciens, and the function of fababean FT gene in plant flowering was identified. The results showed that the full length of FTa1 gene was 216bp, while that of FTa2 gene was 531bp. The overexpression of FTa2 gene can significantly promote flowering in Arabidopsis thaliana, while FTa1 gene can not promote flowering, which may be due to the fact that FTa1 protein is a dysfunctional truncated protein, suggesting the defunctionalization of repetitive genes in the evolution of fababean.
KEY WORDS：fababean；FT homologous gene；cloning；florigen
引言

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