Construction of a new GFP vector and its use for Fusaruim oxysporum transformation

In this study, the gfp fragment as a reporter gene had integrated into the form plasmid vector pBC-hygro which contains an expressive promoter of the fungus to facilitate the transformation of Fusarium oxysporum. The resultant plasmid pBC-hygro-GFP was identified by digestion with enzymes. Binary plasmids pBC-hygro-GFP were transformed into F. oxysporum by using the PEG–CaCl2 mediated transformation technique. Results show that the recombinant plasmid pBC-hygro-GFP was constructed correctly. The gfp gene was stably maintained and did not convey any significant loss of phenotype which would affect the survival and behaviour of the tagged strains. Introduction of the gfp gene into F. oxysporum provides a simple, specific and cost-effective method of strain identification for ecological studies. Transcriptional reporter vectors were constructed for using the green fluorescent protein (GFP) reporter.