| Autor: |
Pérez-Pérez, W. D., Carrasco-Navarro, U., García‑Estrada, C., Kosalková, K., Gutiérrez-Ruíz, M. C., Barrios-González, J., Fierro, F. |
| Rok vydání: |
2022 |
| DOI: |
10.6084/m9.figshare.19500863.v1 |
| Popis: |
Additional file 4. Confirmation of the presence of plasmids for overexpression of Pc-yap1 and Pc-rsmA in P. chrysogenum transformants. (A) Scheme of the fusion of the pki promoter with the Pc-yap1 (top) and Pc-rsmA (down) genes at the ATG start codon in the plasmids pPyrG-pki::Pc-yap1 and pPyrG-pki::Pc-rsmA. Primers Pki1(gpd1)F, OE-PcYap1-R, siYAP1-R, OE-RsmA-R and siRSMA-R are shown at the position of annealing with sequences in the pki promoter and the Pc-yap1 and Pc-rsmA genes, respectively. The expected size of amplified DNA fragments in each type of transformant for every primer pair is indicated with double-headed arrows. (B) Agarose gel with the result of PCR amplification with primers Pki1(gpd1)F and OE-PcYap1-R using as template DNA from the purified pPyrG-pki::Pc-yap1 plasmid (lane P), DNA from strain Wis54-1255 (lane W), a transformant with the empty pBKSpyrG vector (lane Y) and transformants OE::PcYap1-A through –F (lanes A through F). Transformants selected for further characterization are indicated with a red circle. (C) Result of PCR amplification with primers Pki1(gpd1)F and OE-RsmA-R using as template DNA from transformants OE::PcRsmA-A through –H (lanes A through H). (D) Result of PCR amplification with primers Pki1(gpd1)F and siRSMA-R using as template DNA from the purified pPyrG-pki::Pc-rsmA plasmid (lane P), total DNA from strain Wis54-1255 (lane W), a transformant with the empty pBKSpyrG vector (lane Y) and transformants OE::Pc-rsmA-C through –H (lanes C through H). Transformants selected for further characterization are indicated with a red circle. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
