Johnson SM, Kerekes KM, Zimmermann CR, Pappagianis D; American Society for Microbiology. General Meeting.
Abstr Gen Meet Am Soc Microbiol. 1999 May 30-Jun 3; 99: 298 (abstract no. F-16).
UC Davis, CA.
A crude subcellular vaccine derived from formalin killed Coccidioides immitis spherules, designated F27K, has been shown to protect mice against a lethal intranasal challenge. Con A lectin affinity chromatography was useds to separate components of the F27K vaccine. SDS-PAGE of the material eluted yielded multiple discrete bands. One of these bands, 45 kDa was subjected to N-terminal amino acid sequencing, The derived 18 amino acid sequence showed greatest homology to fungal aspartyl proteases. Degenerate oligonucleotide primers derived from the 45 kDa N-terminal sequence and from a consensus region of several aspartyl proteases were pre- pared and were used to amplify using PCR the intervening region of C. immitis cDNA. The 870 bp amplimer was cloned and the nucleotide sequence determined. The deduced amino acid sequence shows 85.9 and 82.4% homology to aspartyl proteases from Aspergillus fumigates and A. niger respectively. These data support the putative identification of the protein as an aspartyl protease. Studies are underway to clone the entire gene and to express the recombinant pro- tein. This will permit evaluation of this protein as a possible immunogen.
Publication Types:
Keywords:
- Amino Acid Sequence
- Animals
- Aspartic Endopeptidases
- Base Sequence
- Coccidioides
- DNA Primers
- DNA, Complementary
- Electrophoresis, Polyacrylamide Gel
- Mice
- Niger
- Polymerase Chain Reaction
- Sequence Homology, Amino Acid
- genetics
Other ID:
UI: 102195644
From Meeting Abstracts