河南农业科学2018，Vol.47Issue(1)：96-102,7.DOI:10.15933/j.cnki.1004-3268.2018.01.017
虎杖转录因子PcMYB1基因的克隆及原核表达
Cloning and Prokaryotic Expression of Transcription Factor PcMYB1 Gene from Polygonum cuspidatum
摘要
Abstract
In order to identify the role of MYB transcription factors in phenylpropanoid pathway of Polygonum cuspidatum,the MYB homologue gene PcMYB1 (GenBank accession No.KY495789) was amplified from the leaf of P.cuspidatum by RT-PCR and RACE methods.The full length cDNA of PcMYB1 is 1 152 bp and the open reading frame is 813 bp,encoding a protein of 270 amino acids with relative molecular mass of 30.455 ku.Sequence analysis indicated that the deduced amino acids had common characteristics of MYB transcription factors with two MYB domains of R2 and R3 at the N-terminal and a transcriptional repression domain C2 motif pdLNL[D/E]LXI[G/S] at the C-terminal.Phylogenetic tree analysis suggested that the encoded protein had the closest genetic relationship with PhMYB4 from Petunia × hybrida.The recombinant prokaryotic expression vector pET28a-PcMYB1 was constructed by inserting the DNA fragment into the prokaryotic expression vector pET-28a,and then transformed into E.coli BL21 (DE3).SDS-PAGE analysis showed that the size of expressed protein was consistent with expectation.In conclusion,the full-length sequence of PcMYB1 gene in P.cuspidatum is obtained,and the prokaryotic expression vector pET28a-PcMYB1 can be used for functional study of PcMYB1 gene.关键词
虎杖/MYB转录因子/PcMYB1/原核表达/苯丙烷代谢/RACEKey words
Polygonum cuspidatum/MYB transcription factor/PcMYB1/Prokaryotic expression/Phenylpropanoid metabolism/RACE分类
农业科技引用本文复制引用
柳忠玉,雷健,李晓筱,刘婵,覃建兵,许锋..虎杖转录因子PcMYB1基因的克隆及原核表达[J].河南农业科学,2018,47(1):96-102,7.基金项目
国家自然科学基金项目(31670608) （31670608）
长江大学湿地生态与农业利用教育部工程研究中心开放基金项目(KF201511) （KF201511）
长江大学校级大学生创新训练计划项目(2017073) （2017073）
