垂丝海棠花色素苷合成基因MhDFR的克隆

Cloning of MhDFR Gene Involved in the Anthocyanin Biosynthesis in Malus Halliana(Voss.) Koehne

目的:利用同源克隆法从垂丝海棠(Malus halliana(Voss.)Koehne.)中克隆到花色素苷合成相关酶DFR(二氢黄酮醇-4-还原酶)基因,这为进一步研究基因表达和基因功能奠定基础。方法:采用CTAB法提取垂丝海棠叶片总RNA,通过RT-PCR克隆得到DFR基因。结果:得到一个长度为1 181 bp的DFR基因,该基因编码394个氨基酸残基,通过数据库进行比对分析,表明实验得到的DFR基因和其他植物中的DFR基因在编码的氨基酸上具有很高的同源性,因此命名为MhDFR,另外MhDFR与观赏海棠"王族"的McDFR在进化上亲缘关系最近近。结论:通过MhDFR的克隆,为进一步研究色素的合成及代谢奠定基础。

Objective:Cloning of MhDFR(Dihydroflavonol 4-reductase) gene involved in the anthocyanin biosynthesis in Malus Halliana(Voss.) Koehne.using homologous method,was the foundation for further gene expression and gene function study.Method:The total RNA was extracted by CTAB method from the leaves of M.Halliana,and DFR gene was cloned by RT-PCR.Result:An 1181 bp gene was obtained,which encoded 394 amino acid residues.This gene showed high homolog to DFR gene family in other plant species when compared against NCBI database,and so it was named MhDFR.Phylogenetic tree of MhDFR demonstrates that MhDFR showed high relationship with McDFR in origin.Conclusion:Therefore,it is the foundation for study anthocyanin biosynthesis and metabolism on the base of cloning of MhDFR in further.