中国现代中药2017，Vol.19Issue(6)：753-757,763,6.DOI:10.13313/j.issn.1673-4890.2017.6.004
雷公藤4α-甲基氧化酶基因(TwSMO1)的cDNA全长克隆及表达分析
Cloning and Expression Analysis of 4α-methyl Oxidase 1 cDNA in Tripterygium wilfordii
摘要
Abstract
Objective:To obtain the full-length cDNA of 4α-methyl oxidase 1(TwSMO1)from Tripterygium wilfordii,and further demonstrate the sequence information and function.Methods:Based on the transcriptome data of T.wilfordii,the rapid-amplification of cDNA ends (RACE) technology was used to clone 4α-methyl oxidase gene (SMO1).The bioinformatics software was used to analyze the information of nucleic acid sequence and the coding amino acids sequence. T.wilfordii suspension cells were induced by methyl jasmonic acid (MeJA),and the genetic differential expression of TwSMO1 were detected through RT-PCR method.Results:The full-length cDNA sequence of TwSMO1 gene was 1505 bp,containing a 900 bp open reading frame and encodes 299 amino acids.The prediction of molecular weight of coding protein was 34.55 kDa and the theory of isoelectric point was 6.89.Multiple sequence alignment results showed that the amino acid sequence of TwSMO1 had a high homology with other species,including three conservative domains of SMO1 gene family.Phylogenetic tree analysis indicated that the gene was classified as first family of 4α-methyl oxidase gene,hence we named it TwSMO1.RT-PCR results showed that,after induced by MeJA,the level of TwSMO1 gene expression was about 450 times higher than the control group at 1 h.Conclusion:The TwSMO1 gene was cloned from T.wilfordii firstly,the bioinformation of the sequence was analyzed and the enzyme function was verified.The results laid the foundation for further research on its gene function and sterol biosynthetic pathway of T.wilfordii.关键词
雷公藤/4α-甲基氧化酶/克隆/生物信息学分析/诱导表达Key words
Tripterygium wilfordii/4α-methyl oxidase/cloning/Bioinformatics analysis/inducible expression引用本文复制引用
关红雨,胡添源,赵瑜君,苏平,童宇茹,张逸风,张夏楠,高伟,黄璐琦..雷公藤4α-甲基氧化酶基因(TwSMO1)的cDNA全长克隆及表达分析[J].中国现代中药,2017,19(6):753-757,763,6.基金项目
国家自然科学优秀青年科学基金项目(81422053) （81422053）
国家杰出青年科学基金项目(81325023) （81325023）
