Expression of the blue and yellow fluorescent proteins in Ganoderma lingzhi and application of the yellow fluorescent protein in activity analysis of inducible promoter Pcbh2
The blue fluorescent protein expression plasmid PJW-EXP-intron-opbfp and yellow fluorescent protein expression plasmid PJW-EXP-intron-opyfp were constructed and transformed into protoplasts of Ganoderma lingzhi.Transformants were selected by PCR and subsequently microscopically screened for fluorescent signals.The pJW-EXP-intron-opbfp and pJW-EXP-intron-opyfp transformants respectively exhibited blue and yellow fluorescence in the mycelia.However,no fluorescence was detected in wild-type(WT)strains.These results showed that blue and yellow fluorescent proteins were successfully expressed in G.lingzhi.Moreover,the activity of the cellulase Ⅱ gene promoter(Pcbh2)was evaluated in G.lingzhi using yfp as a reporter gene.The yfp-Pcbh2 transformants were obtained by genetic transformation of the plasmid yfp-Pcbh2 into protoplasts of G.lignzhi.Under induction of inducer avicel,the mycelia of the yfp-Pcbh2 transformants emitted yellow fluorescence signals,while no fluorescence was detected in the mycelia without the inducer.These results showed that Pcbh2 was successfully activated by avicel in G.lingzhi.
万方数据
维普
