摘要
为了获得金钗石斛(Dendrobium nobile)过氧化物酶基因(命名为DnPOD)编码序列,分析该基因的分子特征,掌握该基因原核表达数据,本研究采用同源克隆的方法,以金钗石斛cDNA为模板,设计基因特异性引物克隆DnPOD基因,分析了DnPOD基因结构,将重组质粒pET28α-DnPOD在BL21(DE3)菌株中诱导表达,构建了DnPOD蛋白3D结构以及进化树。结果表明,DnPOD基因编码区长999 bp,编码332个氨基酸,该基因具有4个外显子和3个内含子。经IPTG诱导表达的蛋白质分子量为37.31 kD。对金钗石斛DnPOD基因克隆和原核表达研究,可为进一步研究该基因的生物学功能打下基础。
In order to obtain the coding sequence of the peroxidase gene(named DnPOD)of Dendrobium nobile,analyze the molecular characteristics and master the prokaryotic expression data,the homologous cloning method was used to design the gene specific primers to isolate the DnPOD gene from D.nobile cDNA.The gene structure was analyzed,and protein 3 D structure and phylogenetic tree of DnPOD proteins were constructed.The recombinant vector pET28α-DnPOD was induced to express in Escherichia coli BL21(DE3)strain.The results showed that the DnPOD gene coding sequence length is 999 bp and encodes 332 amino acids.The gene has 4 exons and 3 introns.The molecular weight of the protein induced by IPTG is about 37.31 kD.The study on cloning and prokaryotic expression of DnPOD gene of D.nobile can lay the foundation for further research on the biological function of this gene.
作者
韩洪强
周培富
吴友发
李兰兰
施家鑫
汤洪敏
Han Hongqiang;Zhou Peifu;Wu Youfa;Li Lanlan;Shi Jiaxin;Tang Hongmin(Cell and Molecular Biology Laboratory,School of Ethnic-MinorityMedicine,Guizhou Minzu University,Guiyang,550025;School of Pharmaceutical Sciences,Guizhou Medical University,Guiyang,550025;Guizhou Bailing Pharmaceutical Co.,Ltd.,Anshun,561000)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2021年第5期2254-2261,共8页
Genomics and Applied Biology
基金
贵州省科技厅联合基金项目(黔科合LH[2015]7209)
贵州民族大学人才引进项目(15XRY013)
贵州省大学生创新创业训练计划项目(201710672005)共同资助
关键词
金钗石斛
过氧化物酶
基因克隆
基因结构
原核表达
Dendrobium nobile
Peroxidase
Gene cloning
Gene structure
Prokaryotic expression
