Expression of egfp gene based on Agrobacterium tumefaciens-mediated transformation in Beauveria bassiana sensu lato ERL836

Abstract

In the present study, an enhanced green fluorescent protein gene (egfp) was expressed in Beauveria bassiana ERL836 based on the Agrobacterium tumefaciens-mediated transformation (AtMT) method. The ERL836 transformants were generated with pCambia-egfp binary vectors. Ten transformants were randomly selected and analyzed for the T-DNA insertion and gene expression. The results revealed that 60% (egfp) of the fungal putative transformants were inserted by the T-DNA fragment. The expressions of egfp in putative transformants were rapidly detected by reverse transcription PCR (RT-PCR) method. Among these transformants, 33.33% (egfp, 2 transformants) expressed the target genes. Two selected egfp transformants showed strong green fluorescence with different expression levels. The results of this study could provide a successful procedure for foreign protein production in B. bassiana by using the AtMT method and accomplished with a rapid selection of fungal transformants using RT-PCR.