Fig. 1
From: Role of Snf-β in lipid accumulation in the high lipid‐producing fungus Mucor circinelloides WJ11
Knockout and overexpression of Snf-β gene a Construction of knockout plasmid pUC1576 (lower). Homologous recombination event between wild type locus (middle) with the replacement fragment (upper). F4, R4, R5 are indicating the primers. The arrows showing the expected PCR band size using the indicated primers. b Verification of knockout transformants with indicated primers (Left side). Lane M: Marker, 1–2: knockout transformants (MU1576 and MU1577 respectively), 3: Wild type control strain (MU2075) using F4 and R4 primers. Lane M: Marker, 2–3: knockout transformants (MU1576 and MU1577), 1: control strain (MU2075) using F4 and R5 primers (Right side). c Plasmid structure of pMAT2075-Snf-β for Snf-β overexpression in M. circinelloides is shown. 2F, 2R, 3F and 3R are indicating the primers. d PCR amplification of Snf-β gene and plasmid region using primers 2F/2R. Lane M: Marker, Lane 2, 3, 4 showing the presence of Snf-β gene in overexpressing transformants Mc3075, Mc3076, Mc3077 respectively and Lane 1 representing the control strain. Verification of the overexpressing strain by amplification of the whole carRP locus with the primers 3F/3R. Lane M: Marker, Lane 2, 3, 4 showing the presence of Snf-β gene including whole carRP locus in overexpressing transformants Mc3075, Mc3076, Mc3077 respectively and Lane 1 representing carRP locus in Mc2075 strain without Snf-β gene (carrying the empty vector pMAT2075). Sizes in the kb of the relevant fragment are indicated