马铃薯StFD2基因的克隆及初步功能分析
毕业论文关键词：马铃薯；FD基因；开花途径
Molecular cloning and primary function analysis of potato StFD2
Abstract：Flowering is an important turning point from vegetative growth to generative growth. The initial of floral pathway is vital to the success of generative growth. In model plant Arabidopsis thaliana, an important florigen protein FLOWERING LOCUS T (FT) interacted with a downstream bZIP protein FLOWERING LOCUS D (FD) and coregulated flowering process in SAM. In potato, the studies of flowering are rare, but it is very vital for sexual breeding. For now, the FT protein SP3D was cloned, but whether it functions with FD on flowering is still elusive. In our studies, we got the FD candidate gene StFD2 through homologous alignment, then we cloned the sequence using genomic DNA and cDNA from different tissues and did the bioinformatic analysis. All these results showed that StFD2 gene has a 684bp-long ORF coding 227 amino acids, and the molecular weight is around 24 kD and pl is 9.71. StFD2, as a non-secreted protein, contained a bZIP domain. PCR assay showed its expression is very low in all tested tissues, and only can be detected in leaves. All these information offers theoretical base to further research of its function in flowering.
Key words: Solanum tuberosum L；FD；floral pathway
目 录
摘要 1
关键词 1
Abstract 1
Key words 1
1 材料与方法 3
1．1 材料 3
1．1．1 植物材料 3
1．1．2 菌株和载体 4
1．1．3 酶与化学试剂 4
1．2 方法 4
1．2．1 基因组DNA和总RNA的提取 4
1．2．2 StFD2序列的扩增 4
1．2．3 DNA片段测序 4
1．2．4 cDNA第一链的逆转录合成 4
1．2．5 组织特异性分析 4
1．2．6 生物信息学分析 5
2 结果与分析 5
2．1 StFD2基因的获得与克隆 5
2．1．1 序列的比对获得 5
2．1．2 基因组片段电泳结果 5
2．1．3 基因组PCR胶回收测序结果（见附录A） 6
2．1．4 NCBI序列比对 6
2．1．5 组织特异性分析 6
2．2 生物信息学分析 6
2．2．1 从Spud DB网站下载得到基因CDS序列（见附录B） 6
2．2．2 蛋白质结构与定位分析 6
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