Reacción #1885014

ord-c106cb2cf04244a8a22f452336890bc3

Ecuación de reacción

O=c1cc[nH]c(=O)[nH]1
uracil
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
OC[C@@H](O)[C@H](O)[C@@H](O)CO
Xylitol

Reactivos

Ninguno

Condiciones de reacción

Temperatura
30°CELSIUS
Condiciones detalladas
See reaction.notes.procedure_details.

Tratamiento posterior

  1. 1
    workup.ADDITIONabove was introduced into Candida parapsilosis in which URA3
  2. 2
    OtroThe colonies formed on the solid medium
  3. 3
    workup.ADDITIONintroduced with the cassette
  4. 4
    ExtracciónThe 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.

Procedimiento

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.

Fuente

DOI: 10.6084/m9.figshare.5104873.v1Patente: US08343736B2uspto-grants-2013_01