Skip to main content
Fig. 6 | Biotechnology for Biofuels

Fig. 6

From: Blocking drug efflux mechanisms facilitate genome engineering process in hypercellulolytic fungus, Penicillium funiculosum NCIM1228

Fig. 6

CRISPR/Cas9-based cbh1 gene deletion in P. funiculosum NCIM1228. a Schematic representation of cbh1 ORF to show the position of sgRNA I and II (highlighted in pink) used for complete removal of cbh1 ORF. 10 hygromycin-resistant transformants were tested for integration of repair template at the target site. Two sets of primers were used; b Amplification of 1319 bp fragment amplified by primers CBH1 1 kb up F and Hph int R. The amplicon refers to the region 1 kb upstream to cbh1 ORF and 200 bp of hph resistance cassette. c Amplification of 1828 bp fragment amplified by primers CBH1 1.5 kb dn R and Hph int F. The amplicon refers to the region 1.5 kb downstream to cbh1 ORF and 200 bp of hph resistance cassette. d Schematic representation of cbh1 ORF and flanking region to show the position of sgRNA III and IV (highlighted in yellow) used for complete removal of cbh1 ORF. 10 hygromycin-resistant transformants were tested for integration of repair template at the target site. e Amplification of 1319 bp fragment amplified by primers CBH1 1 kb up F and Hph int R. f Amplification of 1828 bp fragment amplified by primers CBH1 1.5 kb dn R and Hph int F

Back to article page