Plasmid preparation
The plasmid pANGFPHPH was used in all experiments. It contained
the strong promoter of glyceraldehyde 3-phosphate dehydrogenase
(gpdA) of Aspergillus nidulans. This sequence provided
high levels of expression of the Hygromycin resistance gene
(hph). Fused to this gene the anthranilate synthetase terminator
(trpC) allowed a correct expression of the resistance gene. It also
contained a fragment of the gpdA promoter joined to the GFP sequence and the nopaline synthase terminator. The fragments
mentioned above were commercially synthesized by GenScript
and cloned in the plasmid BlueScript. For bacterial cloning, Escherichia
coli (E. coli) strain DH5a was used. Plasmid extractions were
performed as described by Sambrook and colleagues