Isolation and characterization of an immunogenic fragment of heat shock protein 60 from Trichophyton mentagrophytes.
作者： M. Raska , E. Zemanova , L. Kafkova , J. Belakova , N. K. Vudattu
DOI: 10.1111/J.1439-0507.2004.01046.X
关键词:
摘要: Summary Heat shock protein 60 (hsp60) were isolated from several fungal, protozoal and many bacterial pathogens successfully used for protective vaccination in some infection models. This work focuses on the isolation of recombinant hsp60 dermatophyte, Trichophyton mentagrophytes as a potentially antigen trichophytosis. With help previously tested set degenerated primers, it was reverse transcriptase polymerase chain reaction (RT-PCR) partial cDNA T. (labelled hsp60-TM814), which cloned into cloning vector. The sequencing hsp60-TM814 global alignment confirmed homology with other members family. Hsp60-TM814 corresponds to region encoding immunoprotective fragment Histoplasma capsulatum, mouse model histoplasmosis. A (r-hsp60-TM664), 220 amino acids length, prepared prokaryote expression system, its identity by mass spectroscopy. High immunogenicity r-hsp60-TM664 proven after subcutaneous immunization mice. Immunized sera recognized Western blots well liver lysate Candida albicans. Zusammenfassung Hsp60-Proteine wurden aus verschiedenen pathogenen Pilzen, Protozoen und Bakterien isoliert mit Erfolg fur protektive Vakzination Infektionsmodellen benutzt. In unseren Untersuchungen konzentrierten wir uns auf die Isolierung des rekombinanten hsp60-Proteins dem Fadenpilz als potenziell protektiven Antigens bei der Trichophytose. Mit Hilfe von vorher getesteten Primern benutzten RT-PCR zur einer partialen mentagrophytes-hsp60 (bezeichnet einen Klonierungsvektor kloniert wurde. Sequenzierung hsp60-TM814-cDNA globales Alignment bestatigten Homologie anderen Genen Hsp 60-Familie. entspricht codierenden Region das immunogene, Fragment erfolgreich im Mausemodell Histoplasmose benutzt Ein rekombinantes Aminosauren lang, wurde einem prokaryotischen Expressionssystem synthetisiert seine Identitat durch Massenspektroskopie bestatigt. Subkutane Immunisierung Mausen hat hohe Immunogenitat nachgewiesen. An den Westernblots reagierte Serum immunisierten r-hsp60-TM664-Protein weiter hsp60-Proteinen Lysaten Mauseleber Pilz albicans.
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