Figure 2. Validation of pLUO vector in P. lutzii. (A) Representative image of experiment workflow. The first selection plate contains Cefotaxime 100 μg/mL to kill the remaining A. tumefaciens colonies. Afterwards, fungal colonies are selected on BHI containing only Hygromicin B for three rounds, and then alternating media with or without Hygromicyn until reaching nine rounds of mitotic stability. (B) P. lutzii colonies transformed with pLUO vector after reaching mitotic stability. (C) Eletrophoresis gel showing the colony PCR reactions from A. tumefaciens as a positive control with the hph gene amplified by its primers (resulting in a band of 705 base pairs; the lower band is an unespecific one that always appears for A. tumefaciens). (D) Eletrophoresis gel showing the PCR reactions of P. lutzii transformants. M, molecular ladder; and C, negative control of the reaction.
