Chinese Journal of Dermatology ›› 2019, Vol. 52 ›› Issue (3): 189-192.doi: 10.3760/cma.j.issn.0412-4030.2019.03.008

• Original Articles • Previous Articles     Next Articles

Localization of PQ-LRP protein in Microsporum canis

Liu Yang, Xu Yu, Zhang Furong, Tan Can, Yang Guoling   

  1. Department of Dermatology, The First Affiliated Hospital of Dalian Medical University, Dalian 116011, China
  • Received:2018-06-06 Revised:2018-12-26 Online:2019-03-15 Published:2019-03-04
  • Contact: Yang Guoling E-mail:Yanggl@medmail.com.cn
  • Supported by:
    National Natural Science Foundation of China (81071330)

Abstract: 【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

CLC Number: 

  • R751