Chinese Journal of Dermatology ›› 2010, Vol. 43 ›› Issue (12): 878-879.

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Construction of plasmid vector pSilent-GFP for assessing the gene functions of Penicillium marnefei

  

  • Received:2010-04-06 Revised:2010-05-17 Online:2010-12-15 Published:2010-12-13

Abstract:

Objective To develop a new tool to study the gene function of Penicillium marneffei based on pSilent-1 vector. Methods GFP gene was subcloned into the pSilent-1vector and identified by sequence analysis, then, the reconstructed plasmid pSilent-GFP was transformed into the yeast cells of Penicillium marneffei by electronic transformation. The positive clones were selected by hygromycin and by directed PCR to amplify the GFP gene. The expression of GFP could be detected under inflorescent microscopy. Results It was confirmed that gene of GFP was successfully reconstructed into pSilent-1 plasmid and obtained the pSilent-GFP plamid. The pSilent-GFP plasmid could be transformed into Penicillium marneffei and expressed the GFP in yeast and mycelia phases of Penicillium marneffei. Conclusions The pSilent-GFP could stably express in both phases of, and could be used as a tool to study the gene function of Penicllium marneffei.