Reaction #1885014

Reaction equation

uracil

glucose

→

Xylitol

Conditions

Workup

1. 1
   workup.ADDITIONabove was introduced into Candida parapsilosis in which URA3
2. 2
   OtherThe colonies formed on the solid medium
3. 3
   workup.ADDITIONintroduced with the cassette
4. 4
   ExtractionThe colonies were inoculated in 4 ml of YM medium (glucose 20 g/L, yeast extract 3 g/L, malt extract 3 g/L, and peptone 5 g/L)
5. 5
   workup.WAITfollowed by incubation for 2 days at 30° C.

Procedure

The expression cassette PGtrpfs2-CoAraA constructed above was introduced into Candida parapsilosis in which URA3 was inactivated, which was smeared on the uracil-free solid selection medium (yeast nitrogen base 6.7 g/L, glucose 20 g/L, and agarose powder 15 g/L), followed by incubation for 2 days at 30° C. The colonies formed on the solid medium were the strains introduced with the cassette. The colonies were inoculated in 4 ml of YM medium (glucose 20 g/L, yeast extract 3 g/L, malt extract 3 g/L, and peptone 5 g/L), followed by shaking-culture at 150 rpm for 12 hours at 30° C. Then, the culture solution was smeared on 5-FOA solid medium (yeast nitrogen base 6.7 g/L, glucose 20 g/L, 5-FOA 0.8 g/L, uracil 0.1 g/L, and agarose powder 15 g/L), followed by incubation for 2 days at 30° C.

Source