玉米大斑病菌转录因子StSte12的活性及其对酵母生长的功能互补分析

Transcription Activity of a Transcription Factor StSte12 from Setosphaeria turcica and Function Analysis Through Yeast Complementation

【目的】分析玉米大斑病菌StSte12的结构、转录活性及功能。【方法】利用生物信息学方法,对获得的StSte12进行保守结构域和进化树分析,推测该基因的功能;利用β-半乳糖苷酶法检测StSte12的转录活性;将StSTE12转化至酿酒酵母ScSTE12基因缺失突变体ste12Δ中,筛选酿酒酵母ScSTE12的功能互补突变体并对其分析,验证玉米大斑病菌StSTE12的功能。【结果】通过蛋白比对发现StSte12具有转录因子特有的STE homeodomain和ZnF_C2H2锌指结构;同源性分析显示,该基因与其它植物病原真菌的STE12-like基因有较高的同源性;利用β-半乳糖苷酶法检测发现,StSte12具有转录激活活性;酵母互补试验表明,StSTE12可以回复酿酒酵母ste12Δ的功能,能够调控酵母细胞的生长。【结论】玉米大斑病菌StSTE12属于STE12-like基因;转录因子StSte12具有转录活性;对酵母细胞在YPD培养基上的侵入生长有重要的调控作用。

[ Objective ] The objective of this study is to identify the structure characteristics, transcription activity and function ofStStel2 fromSetosphaeriaturcica. [Method]Conservative domain prediction andphylogenetic tree analysis were used to predict the possible function of StSte 12 through bioinformatics analysis. The transcription activity of StSTE12 was tested by fl-galactosidase activity method. Through characterization of growth of the StSTE12 complement transformant, in which StSTE12 was introduced into ScSTE12 null mutant of Saccharomyces ceresivisiae, the function of StSTE12 was preliminarily identified. [ Result ] StSte 12 contained STE homeodomain and ZnF_C2H2 structure, which was the characteristics of Stel2-1ike transcription factor. Sequence analysis revealed that StSTE12 shared highly homology with other STE12-1ike genes of plant pathogens. StStel2 had transcription activation in vitro and could restore the function ofstel2A ofS. ceresivisiae, which regulated invasion growth of the yeast cell. [ Conclusion] Transcription factor StSTE12, a STE12-1ike gene from S. turcica, has a transcription activity and plays an important role in regulating the invasion growth of the cell on YPD medium.