构建fks1基因缺陷型酿酒酵母基因工程菌改良成膜能力

Improvement of Saccharomyces cerevisiae biofilm forming ability by genetically constructing fks1 mutant

β-1,3-葡聚糖作为致病酵母菌生物膜的组成成分,在抗药性方面发挥着重要作用。为了将生物膜应用于工业化酿酒酵母,有必要了解其对应的膜成分及成膜机制。本文中,笔者旨在研究编码β-1,3-葡聚糖酶催化亚基的fks1基因缺失对酿酒酵母成膜能力的影响,以期了解酿酒酵母成膜机制。利用同源重组原理,将抗性基因片段替换插入到目的基因片段中,使fks1基因失活,然后通过96孔板结晶紫染色、平板入侵、扫描电镜成像等表型实验验证其成膜能力。结果表明:当fks1基因缺失时,酿酒酵母1308的成膜能力增强,这为生物膜的下一步应用提供了理论依据。

As a component of biofilm in pathogenic yeast,β-1,3-glucan plays an important role in resistance to drugs.In order to the application of industrial yeast’s biofilm,it is necessary to study its mechanism and component.We studied the impact on the formation of yeast biofilm in the absence of fks1 gene,and provide basis for the improvement of biofilm in the future.Based on the principle of homologous recombination,the resistant gene fragment was inserted into the target gene fragment to inactivate the fks1 gene.Then we tested the ability of forming biofilm by crystal violet staining in 96-well plate,flocculation,invasion in agar plate,and image through scanning electron microscope(SEM).The biofilm forming ability of Saccharomyces cerevisiae 1308 was enhanced in the absence of fks1 gene.This provides a theoretical foundation for the application of biofilm.