Molecular cloning and expression of a cDNA encoding a Coccidioides posadasii Cu,Zn superoxide dismutase identified by proteomic analysis of the coccidioidal T27K Vaccine

Jennine M. Lunetta, Keira A. Simmons, Suzanne M. Johnson, Demosthenes Pappagianis

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Scopus citations

Abstract

Previous studies have demonstrated that the coccidioidal T27K vaccine preparation is protective in mice against respiratory challenge using Coccidioides posadasii (C. posadasii) arthroconidia. Proteomic methods have been employed to define the molecular components within the vaccine. This method has led to the identification of novel and previously uncharacterized coccidioidal proteins including a Cu,Zn superoxide dismutase. A two-dimensional gel of the T27K vaccine was run and spots were excised for mass spectrometric analysis. One peptide was obtained from the T27K gel that matched a TIGR C. posadasii 2.0 gene index tentative consensus sequence, TC1072, which is similar to fungal Cu,Zn superoxide dismutase. Activity assays performed with native PAGE gels of the T27K vaccine showed that the vaccine contains superoxide dismutase. The cDNA encoding the enzyme has been cloned and sequenced and expressed as a recombinant protein.

Original languageEnglish (US)
Title of host publicationAnnals of the New York Academy of Sciences
Pages181-197
Number of pages17
Volume1111
DOIs
StatePublished - Sep 2007

Publication series

NameAnnals of the New York Academy of Sciences
Volume1111
ISSN (Print)00778923
ISSN (Electronic)17496632

Keywords

  • Coccidioides
  • Fungus
  • Mass spectrometry
  • Metalloenzyme
  • Rapid amplification of cDNA ends
  • Reverse-transcription polymerase chain reaction
  • SOD1
  • Superoxide dismutase
  • Two-dimensional gel electrophoresis
  • Vaccine

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Fingerprint Dive into the research topics of 'Molecular cloning and expression of a cDNA encoding a Coccidioides posadasii Cu,Zn superoxide dismutase identified by proteomic analysis of the coccidioidal T27K Vaccine'. Together they form a unique fingerprint.

Cite this