中药材2025,Vol.48Issue(10):2407-2413,7.DOI:10.13863/j.issn1001-4454.2025.10.001
桑黄转录因子基因SbMYB1的克隆、表达及转化酵母抗胁迫分析
Cloning,Expression and Transformation of Transcription Factor Gene SbMYB1 from Sanghuangporus baumii into Saccharomyces cerevisiae for Stress Resistance Analysis
摘要
Abstract
Objective:To clone,expresse and transform the transcription factor gene SbMYB1 from Sanghuangporus baumii into Saccharomyces cerevisiae and conduct the stress resistance analysis on transformed Saccharomyces cerevisiae,laying the foundation for re-vealing the function of the Sanghuangporus baumii transcription factor gene SbMYB1.Methods:The Sanghuangporus baumii transcrip-tion factor gene SbMYB1 was cloned and bioinformatics analysis was conducted.The gene expression pattern was detected by real-time fluorescence quantitative PCR.The overexpression vector of the SbMYB1 gene was constructed and genetically transformed into Saccha-romyces cerevisiae,and the growth status of transformed Saccharomyces cerevisiae after abiotic stress treatment was observed.Results:The total length of the SbMYB1 gene from Sanghuangporus baumii was 2 211 bp,containing 6 exons and 5 introns.The total length of the cDNA was 1 929 bp,encoding 642 amino acids.It did not have a signal peptide or transmembrane structure,and the subcellular location was in the nucleus.Phylogenetic analysis indicated that the genetic relationship of SbMYB1 protein was closely related to Fomitiporia mediterranea.The analysis of the cis-type elements of the promoter revealed that SbMYB1 contained functional elements that respond to abiotic stress.qRT-PCR analysis indicated that the SbMYB1 gene responded to cold,heat and heavy metal stress to varying degrees.Overexpression of the SbMYB1 gene from Sanghuangporus baumii enhanced the tolerance of recombinant Saccharomyces cerevisiae to high temperature and heavy metal stress.Conclusion:The SbMYB1 gene from Sanghuangporus baumii is successfully cloned,and it is found that the expression of this gene is significantly induced under abiotic stress.Through the heterologous expression experiment of Saccharomyces cerevisiae,it is confirmed that the SbMYB1 gene from Sanghuangporus baumii has a potentially important function in the response to adverse conditions.关键词
桑黄/MYB转录因子基因/酵母/载体构建/非生物胁迫Key words
Sanghuangporus baumii(Pilát)L.W.Zhou&Y.C.Dai/MYB transcription factor gene/Saccharomyces cerevisiae/Vector construction/Abiotic stress分类
医药卫生引用本文复制引用
孙婷婷,刘瑞鹏,张林芳,杜鹏禹,李亚伟,邹莉..桑黄转录因子基因SbMYB1的克隆、表达及转化酵母抗胁迫分析[J].中药材,2025,48(10):2407-2413,7.基金项目
黑龙江省自然科学基金联合引导项目(LH2022C054) (LH2022C054)