金线兰黄酮相关基因的克隆与生物信息学分析

Cloning and Bioinformatics Analysis of Anoectochilus roxburghii Flavonoid Related Gene

为了对金线兰查尔酮合成酶CHS和黄酮醇合成酶FLS进行基因的克隆和生物学信息分析,本研究参照测序得到的金线兰CHS与FLS基因的序列,设计特异性引物进行扩增。利用软件,对金线兰CHS和FLS蛋白的生物信息进行预测分析。结果显示,金线兰CHS基因的CDS总长度1 185 bp,编码394个氨基酸,金线兰FLS基因的CDS总长度1 035 bp,编码344个氨基酸。金线兰CHS蛋白与FLS蛋白皆是酸性、亲水性、无信号肽、非分泌、非跨膜蛋白,亚细胞定位在其他细胞器上,丝氨酸磷酸化位点具有调控二者构象变化的重要作用。在Blast上进行序列比对,发现金线兰CHS序列和FLS蛋白序列与一些兰科植物的相似度很高,前者序列同源性均高达85%以上,后者同源性高达82%和81%,构建的进化树显示了他们较近的亲缘关系。推测得到的CHS序列即为金线兰查尔酮合成酶的编码序列,FLS蛋白序列即为金线兰黄酮醇合成酶的编码序列。本研究为进一步研究金线兰中黄酮相关基因及其调控提供一定参考。

In order to cloning and bioinformatics analysis of chalcone synthase CHS and flavonol synthase FLS from Anoectochilus roxburghii.In this study,the specific primers were designed for amplification by referring to the sequence of Anoectochilus roxburghii CHS and FLS genes obtained by sequencing.The bioinformatics of CHS and FLS proteins were analyzed and predicted by software.The total length of the CDS of the Anoectochilus roxburghii CHS gene is 1185 bp,encoding 394 amino acids.The total length of the CDS of the Anoectochilus roxburghii FLS gene is 1035 bp,encoding 344 amino acids.Both the CHS protein and the FLS protein of Anoectochilus roxburghii are acidic,hydrophilic,signal-free peptides,non-secretory,non-transmembrane proteins,and subcellular localization on other organelles.Serine phosphorylation sites have an important role in regulating the conformational changes of both.Sequence alignment on Blast showed that the CHS and FLS sequences of Anoectochilus roxburghii were highly similar to those of some Orchidaceae plants.The homology of the former was more than 85%,while that of the latter was 82%and 81%.The phylogenetic tree constructed showed their close relationship.The obtained CHS sequence is th e coding sequence of chalcone synthase and the FLS protein sequence is the coding sequence of flavonol synthase from Anoectochilus roxburghii.This study provides a reference for further study of flavonoid-related genes and their regulation in Anoectochilus roxburghii.