水曲柳节律相关GIGANTEA基因的克隆及同源性比对分析

Cloning and Homology Analysis of Biological Rhythms Related GIGANTEA Gene in Fraxinus mandshurica

[目的]对水曲柳节律相关GIGANTEA基因的克隆及同源性进行比对分析。[方法]根据GenBank中GI基因保守结构域设计简并引物,进行PCR分析,并与其他物种的氨基酸序列进行同源性比对分析。[结果]获得了编码区780 bp的部分序列,可以编码260个氨基酸,获得的GI基因命名为FmGI,与多个物种GI基因有着较高的同源性,其中与葡萄、大豆、苜蓿、毛果杨、蓖麻、黄瓜、菊花、大麦、玉米、小麦、拟南芥、黑麦草、洋葱和云杉等多个物种具有较高一致性。系统发育树分析结果表明,GI基因按照单子叶植物、双子叶植物和裸子植物的进化关系聚为3类,明确了水曲柳GI基因的系统进化关系。[结论]该方法对水曲柳生物节律钟GI基因编码区进行了克隆,并对获得的部分GI蛋白序列进行了同源性比对分析,建立了系统进化树,为进一步获得水曲柳GI基因全长及研究其功能起到一定参考作用。

[ Objective ] To comparatively analyze cloning and homology of biological rhythms related GIGANTEA gene in Fraxinus mandshurica. [ Method] Using degenerate primers corresponding to conserved domains of GI gene in GenBank, PCR analysis was conducted, as well as homology analysis with amino acid sequence of other species. [ Result ] The length of the fragment obtained was 780bp, ORF predicted the protein was component of 260 amino acids. The homology analysis, of the obtained fragment showed that, it had a high consistency with Vitis vinifera, Glycine max, Medicago truncatula, Populus trichocarpa, Ricinus communis, Cucamis sativu, Chrysanthemum seticuspe, Hordeum vulgare, Zea mays, Triticum aestivum, Arabidopsis thaliana, Lolium perenn, Allium cepaand, and Picea abies. Phylogenetic analysis showed that according to monocots, dicots and gymnosperms, the evolutionary relationship of GI gene clustered into three categories. The obtained GI gene clustered with dicots, it showed a clear phylogenetic relationship of GI gene in Fraxinus mandshurica. [ Conclusion] The study provide a reference for further obtaining Fraxinus mandshurica GI gene full length and researching its functions.