CDC42基因的激活和抑制对烟曲霉极性生长影响的初步探究OACSTPCD
The preliminary study on the effect of activation and inhibition of CDC42 gene on polar growth of Aspergil-lus fumigatus
目的 构建烟曲霉CDC42基因显性激活性(dominant active,DA)和显性抑制性(dominant negative,DN)点突变菌株,初步了解这两种点突变对烟曲霉极性生长以及RAC1基因表达的影响.方法 运用分子生物学方法构建CDC42基因激活性(CDC42G14v)和抑制性(CDC42T19N)菌株;光学显微镜观察点突变菌落特征;RT-PCR方法检测Ku80、DA Cdc42和DN Cdc42菌株中RAC1的表达量.结果 成功构建烟曲霉CDC42显性激活性(DA)和显性抑制性(DN)点突变菌株并经测序确认;烟曲霉DA Cdc42和DN Cdc42菌株的菌落生长直径、形成分生孢子数和各时间点孢子发芽率较对照株Ku80都明显降低,且差异均有统计学意义(P＜0.05);RT-PCR结果表明DA Cdc42和DN Cdc42菌株中的RAC1的表达量均较对照株Ku80有所上升(P＜0.05).结论 CDC42的激活和抑制都会对烟曲霉的极性生长产生负面影响;且会影响RAC1基因的表达.
Objective To construct CDC42 gene dominant activation and dominant inhibition point mutant strains and to explore the effects of the two point mutations on the polar growth and RAC1 gene expression of Aspergillus fumigatus.Methods CDC42 gene activation(CDC42G14V)and inhibition(CDC42T19N)strains were constructed by molecular biological methods,the characteristics of point mutation colonies were observed under light microscope,and the expression of RAC1 in Ku80,DA Cdc42 and DN Cdc42 strains was detected by RT-PCR.Results Point mutant strains of CDC42G14V and CDC42T19N were successfully constructed and further confirmed by sequencing.The colony growth diameters,conidia num-bers and spore germination rates of DA Cdc42 and DN Cdc42 were significantly different from those of the control strain Ku80(P＜0.05).The results of RT-PCR showed that the expressions of RAC1 in DA Cdc42 and DN Cdc42 strains were higher than that in the control strain Ku80(P＜0.05).Conclusions The activation and inhibition of CDC42 have negative effects on the polar growth of A.fumigatus and affect the expression of RAC1 gene related with polarity growth.
张梦楠;于婉婉;马彦
山西医科大学第二医院皮肤性病科,太原 030000
基础医学
烟曲霉Cdc42点突变极性生长
Aspergillus fumigatusCdc42point mutationpolar growth
《中国真菌学杂志》 2024 (001)
1-7 / 7
2019年度山西省高等学校优秀成果培育项目(2019KJ025);山西省基础研究计划自然科学研究面上项目(202103021224416)
评论