杜仲MVA途径相关基因全长cDNA序列特征研究

Study on full length cDNA of MVA pathway related genes in Eucommia ulmoides

杜仲幼果和成熟果转录组测序的基础上,分离鉴定了幼果和成熟果MVA途径ACOT、HMGS、HMGR基因全长cDNA序列,并对基因的核苷酸序列及其相应氨基酸序列的组成成分、跨膜结构域、疏水性/亲水性、蛋白质二、三级结构、分子系统进化等进行分析。EuACOT基因全长cDNA为1 248 bp,编码416个氨基酸,属于稳定疏水性蛋白,无明显的跨膜区,在进化上分属于不同的分类群,与烟草和番茄的ACOT蛋白间的亲缘关系最为接近;EuHMGS基因全长cDNA为1 356 bp,编码452个氨基酸,属于不稳定的疏水性蛋白,无明显的跨膜区,在进化上分属于不同的分类群,与茶树的HMGS蛋白间的亲缘关系最为接近;EuHMGR基因全长cDNA为1 770 bp,编码590个氨基酸,属于不稳定的疏水性蛋白,有两个较明显的跨膜区,在进化上分属于不同的分类群,与杜仲的HMGR蛋白间的亲缘关系最为接近;以上三个蛋白的二级结构均是以α-螺旋和螺环结构为主的混合型结构。

This analysis was based on data of the transcriptome sequencing of Eucommia young fruit and mature fruit. The terpene synthesis of MVA pathway involving of 30 Unigene. We identified 3 full-length cDNA in MVA pathway of young fruit and mature fruit, These full-length cDNA were analyzed and predicted by the tools of bioinformatics in the following aspects： composition of the nucleotide sequence and corresponding amino acid sequence, transmembrane structure, hydrophobicity or hydrophilicity, secondary and tertiary structure of protein, molecular phylogenetic evolution and so on.The results are as follows： Full-length cDNA of EuACOT gene has 1 248 bp, and coded 416 aa, EuACOT is stable hydrophobic protein without transmembrane domain, and belong to different taxa in evolution, high consistency with the Nicotiana tabacum and Lycopersicon esculentum in ACOT protein sequences and high structure. Full-length cDNA of EuHMGS gene has 1 356 bp, and coded 452 aa, EuHMGS is unstable hydrophobic proteins without transmembrane domain, and high consistency with the Camellia sinensis in HMGR protein sequences and high structure. Full-length cDNA of EuHMGR gene has 1 770 bp, and coded 590 aa, EuHMGR is unstable hydrophobic proteins with two transmemhrane domanin, and high consistency with the Eucommia ulmoides in HMGR protein sequences and high structure. The secondary structures of above three proteins are predominantly hybrid structure of alpha helical and spiral ring structure and belong to different taxa in evolution.