3.1.4.3. Fluorescence detection of green fluorescent protein
marking transformants.Fluorescence expression of stable
transformants was observed under fluorescence microscope, with results being that the three transformants had a stronger fluorescence expression and some had a weaker expression. For transformants with hygromycin resistance, a small amount of hyphae and spore were selected to be observed under fluorescence microscope. It was discovered that GFP had stronger expression in young hyphae and weaker expression in aging hypha, while the expression was weak in conidium.
3.1.4.4. GFP transformants PCR confirmation. Four transformants having fluorescence properties strongly and two wild strains being genomous, were abstracted to prove that hygromycin-resistant transformants contain the inserted gfp gene. As a template, the application of amplified gfp fragment prim-ers. GFPup1 and GFPdown1 of the PCR amplification results showed that the four transformants contain gfp gene, but the other two wild strains do not contain gfp gene.