DNA was transferred as described above, and each time 1μg trp 1+ plasmid was used for selection, and 1 μ g GPYSK sequence was used.

The transformed protoplasts were inoculated on reclaimed rubber and supplemented with 0.5 mm abts [2,2'-diazo-trans (3- ethylbenzothiazole -6- sulfonic acid]. Transformants were selected and cultured on minimal medium.

The selected transformants were inoculated in the center of YMGT/T gel containing 0.5 mM ABTS and cultured at room temperature. ABTS oxidation mediated by polyphenol oxidase can mark the daily growth of plaque. In order to determine the enzyme activity in the liquid medium (120 rpm, 37℃) cultured in a shaker, the transformant was cultured in a 500ml conical flask and added with 100ml YMG/T liquid medium. In some experiments, 0. 1 and 1.0mM copper sulfate were added to the culture medium respectively. Then, 7 ml of suspended hyphae pre-cultured in sterile YMG/T glue for 5 days were added to the culture medium.

I admit I'm bored. . .