铁皮石斛DoSAMDC-like基因的克隆及原核表达

本研究从药用植物铁皮石斛中克隆得到S-腺苷甲硫氨酸脱羧酶(S-adenosyl-L-methionine decarboxylase)基因(DoSAMDC-like)并进行序列分析和原核表达。SAMDC基因家族具有多个成员,本研究采用同源克隆的方法对铁皮石斛中尚未报道的新成员DoSAMDC-like基因进行克隆,并与拟南芥SAMDC基因家族成员和铁皮石斛基因组注释序列进行比对分析、蛋白质特征及原核表达等分析。基因序列分析表明,DoSAMDC-like基因编码区全长1104 bp,推测编码368个氨基酸,氨基酸序列具有SAM_decarbox超家族共同的保守结构域。多序列比对及进化树结果表明,克隆获得的DoSAMDC-like基因序列对应于基因组测序注释的XM_020825443.2序列,一致性为99.55%,与铁皮石斛SAMDC1基因一致性为80.16%。异源表达结果表明,DoSAMDC-like基因受IPTG诱导,蛋白分子量约为47 kD。DoSAMDC-like基因的克隆和原核表达,可为参与铁皮石斛多胺代谢、生物碱合成相关基因的蛋白质生化活性等研究提供了帮助。

Cloning and Prokaryotic Expression of DoSAMDC-like Gene in Dendrobium officinale

The S-adenosyl-L-methionine decarboxylase like gene(DoSAMDC-like)was cloned from medicinal plant Dendrobium officinale and analyzed by bioinformatics and prokaryotic expression in the study.The SAMDC gene family consists of multiple members in plants.In this study,homology-based cloning was used to clone the new member DoSAMDC-like gene in D.officinale.And the DoSAMDC-like gene was compared with Arabidopsis SAMDC genes and the annotated sequences of D.officinale from the genome sequencing.Multiple alignments,protein characteristics and prokaryotic expression were carried out to analysis the DoSAMDC-like gene.Gene sequence analysis showed that DoSAMDC-like gene CDS length is 1104 bp and encodes 368 amino acids with SAM_decarbox superfamily conserved domain.Multiple sequence alignment and phylogenetic tree results showed that the DoSAMDC-like gene sequence corresponds to XM_020825443.2 from NCBI with 99.55%identity,but 80.16%identity with the DoSAMDC1 gene.The heterologous expression results showed that the DoSAMDC-like gene was successfully induced by IPTG and the molecular weight was about 47 kD.The cloning and prokaryotic expression of DoSAMDC-like gene can offer help for research of polyamine metabolism and protein biochemical activity of alkaloid synthesis in D.officinale.