Characteristics Analysis,Prokaryotic Expression and Product Purifi-cation of Trichoderma asperellum Myb27 Transcription Factor
Using cDNA of the total RNA reverse transcription from the mycelium of Trichoderma CGMCC11653 strain as a template,the full-length 717 bp transcription factor Myb27 coding se-quence was cloned,encoding 238 amino acids polypeptide.The bioinformatics software was used to predict the physical and chemical properties,structural domains and protein subcellular localization,and to analyze the multi-sequence alignment of amino acids and the evolutionary relationship.The re-sults showed that the protein belonged to Myb-like DNA-binding domain and the subcell was lo-cated in the nucleus with a molecular weight of 27 000.The expression of Myb27 changed signifi-cantly under the induction of Alternaria toxin,which may be involved in the positive regulation of the response of Trichoderma against Alternaria toxin stress.The prokaryotic expression vector,pMAL-Emyb27,was constructed,and the transformant ER2523-Emyb27 was obtained by transforming E.coli ER2523.The optimal conditions for successful induction of ER2523-Emyb27 were 0.1 mmol/L IPTG 37 ℃ for 3 h.A single soluble 69 500 recombinant rMyb27 protein was obtained by eluting the MBP label with PBS+10 mmol/L maltose chromatography column.This study has provided recombi-nant proteins in vitro for the cis-acting elements bound by Myb27 transcription factor to regulate the expression of disease resistance genes,and theoretical basis for the mechanism of regulating Tricho-derma against toxin stress of Alternaria alternata.
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维普
