丹参功能基因组学研究Ⅲ——丹参金属硫蛋白MT-2基因的分析

Functional genomics studies of Salvia miltiorrhiza Ⅲ.analyze of metallothionein(MT-2) genes

目的:获得丹参金属硫蛋白基因序列,并进行生物信息学分析和初步的表达特性研究。方法:利用cDNA芯片技术,从不同时期毛状根中获得目的基因。利用BLAST进行序列比对,ORF finder寻找开放读码框,Pro-site分析蛋白质的基本结构域。用半定量RT-PCR检测其在丹参组培苗根、茎、叶中的表达情况。结果:得到2个MT基因全长序列,分别编码80和79个氨基酸,具有Type 2型金属硫蛋白的典型特征,分别命名为SmMT-2 a和SmMT-2b,两者具有71.25%的同源性。半定量RT-PCR表明,MT基因在丹参组培苗根、茎、叶中均有转录水平的表达,叶中表达量高于根和茎。结论:首次得到丹参金属硫蛋白基因,为其功能研究提供了基础。

Objective： To study the metallothionein genes of Salvia miltiorrhiza through bioinformatics and characterization of tissue expression in regenerated shoots. Method： Metallothionein genes were obtained by cDNA microarray analyze. BLAST was used for align, ORF finder software was used to find open reading frame, Prosite database was used to analyze the protein, semi-quantitative RT- PCR method was used to detect the gene expression level. Result： Two metallothionein genes were obtained which were contained a deduced amino acid sequence of 80 and 79 residues, named as SmMT-2a and SmMT-2b, they had a homology of 71.25%. Semi-quantitative RT- PCR indicated that metallothionein genes were expressed in all tissues such as root, stem and leaf in regenerated shoots, while the expression level was higher in leaf than in root and stem. Conclusion： It was the first time that metallothionein genes were obtained from S. miltiorrhiza. It provides a good basis for further functional study of S. miltiorrhiza.