华北农学报2017，Vol.32Issue(3)：65-69,5.DOI:10.7668/hbnxb.2017.03.010
萼脊兰MADS-box基因的克隆及表达载体构建
Cloning and Construction of Expression Vector of MADS-box Gene from Sedirea japonica
摘要
Abstract
In order to study the B type PI gene of "ABC" model, and explore the gene characteristics of petals and stamen formation.In the study,using CTAB approach to extract the total RNA of the petal of Sedirea japonica,and cloning the part-length cDNA sequences of PI gene of Sedirea japonica by using RT-PCR approach.It found that the length of this gene sequence was 633 bp,it encoded 210 putative amino acid residues and the gene sequence had a high homology with Phalaenopsis equestris after sequence alignment.After analysis of the structure and the hydrophilic and the secondary structure of PI protein,it found that the gene sequence contained the MADS-box domain and K-box,it belonged to the MADS-box super family,PI was hydrophilic protein,and it contained 56.19% Alpha helix,13.81% extended strand and 30% random coil.The expression was analysed by Real-time fluorescent quantitative PCR,PI genes mainly expressed in floral organ,and high expression and PI gene expression had tissue specificity.During the construction of expression vector,it leads the sequence containing objective gene and promoter to the pCAMBIA1301 expression vector,it showed that the construction of expression vector 1301-PI was succeed,which laied the fundation for getting new varieties of Sedirea japonica.关键词
萼脊兰/PI基因/克隆/表达分析/表达载体Key words
Sedirea japonica/PI gene/Cloning/Expression analysis/Expression vector分类
农业科技引用本文复制引用
蒋素华,黄萍,王默霏,梁芳,许申平,崔波..萼脊兰MADS-box基因的克隆及表达载体构建[J].华北农学报,2017,32(3):65-69,5.基金项目
郑州市科技发展计划项目(20150448) （20150448）
郑州市重大科技专项(141PZDZX038) （141PZDZX038）
河南省教育厅重点科研项目(14B180036) （14B180036）
