Construction of Bem46 fluorescent localization strain and investigation of the effect on polar growth gene in Aspergillus fumigatus
MA Yan1,CAO Xue1,LI Wen2,LI Xue?jing1,JI Ying1,YANG Jing1,CEN Wen1,FENG Wen?li1
1.Department of Dermatology,The Second Hospital of Shanxi Medical University,Taiyuan 030001,China; 2.Department of Dermatology, Taiyuan Central Hospital, Taiyuan 030009, China
Abstract：Aspergillus fumigatus is an environmental filamentous fungus that can cause life?threatening disease in immunocompromised individuals. BEM46 proteins are evolutionarily conserved, but their functions remain elusive. We constructed a localization strain of Bem46 gene in A. fumigatus with eGFP as the reporter gene was constructed and the distribution of green fluorescent protein in different morphology of A. fumigatus was observed. The expression of Bem1 and Bud5 gene related with polar growth between ΔBem46 and Ku80 was also compared. The Bem46 gene of A. fumigatus was cloned into plasmid pUCGH by molecular biological method and the vector of fluorescent localization strain was constructed. The six transformations of Bem46 localization strains were obtained by the protoplast transformation method and were identified by different PCR. The fluorescence was diffused in the spore and gathered on one side of the spore when the spore was about to germinate by laser confocal microscopy. With the formation of the germ tube, the fluorescence extended on the polar growth side and gradually developed into hypha, and then distributed in the whole hypha. The obvious fluorescence accumulation was found on the tips of the hypha branch. The expression of Bem1 and Bud5 gene in ΔBem46 was lower than the control strain by RT?PCR. In conclusion, it was speculated that Bem46 gene of A. fumigatus may be related to the polar growth of spore budding and the tips of lateral branches of mycelium. The defection of Bem46 gene may affect the expression of Bem1 and Bud5 genes.
马彦, 曹雪, 李雯, 李雪静, 冀英, 杨静, 岑雯, 冯文莉. 烟曲霉Bem46基因荧光定位菌株构建及对极性生长相关基因影响作用初探[J]. 中国人兽共患病学报, 2020, 36(10): 840-846.
MA Yan,CAO Xue,LI Wen,LI Xue?jing,JI Ying,YANG Jing,CEN Wen,FENG Wen?li. Construction of Bem46 fluorescent localization strain and investigation of the effect on polar growth gene in Aspergillus fumigatus. Chinese Journal of Zoonoses, 2020, 36(10): 840-846.
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2020, Vol. 36 
