Abstract
Objective To determine the location of PQ-LRP protein in Microsporum canis using the enhanced green fluorescent protein (EGFP) as a marker. Methods The total RNA was extracted from Microsporum canis, and reversely transcribed into cDNA. The PQ-LRP gene was amplified by PCR using the above cDNA as the template. The fusion gene of PQ-LRP gene and EGFP gene was linked to the plasmid pCAMBIA 1300. Microsporum canis was subjected to Agrobacterium tumefaciens-mediated transformation, in order to achieve the integrated expression of the fusion gene LRP-EGFP in Microsporum canis under the regulation by the fungal universal promoter Ptrpc and terminator Ttrpc. Laser-scanning confocal microscopy was conducted to determine the cellular localization of the fusion protein. Results The expression vector pCAMBIA-LRP-EGFP was successfully constructed, and the fusion gene LRP-EGFP was expressed integratedly in Microsporum canis. Laser-scanning confocal microscopy showed that fluorescence signals of LRP-EGFP were concentrated on the cell membrane of Microsporum canis, giving a granular or cluster-like appearance. Conclusion The infusion gene LRP-EGFP can be successfully expressed in Microsporum canis, and PQ-LRP protein is located on the cell membrane of Microsporum canis. Key words: Microsporum; Fiducial markers; Green fluorescent proteins; Microsporum canis; PQ-LRP; Protein localization
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