Cloning and expression of the complement fixation antigen-chitinase of Coccidioides immitis.

Infection and Immunity
C R ZimmermannD Pappagianis

Abstract

A chitinase had been isolated from the culture filtrates of Coccidioides immitis endosporulating spherules and from hyphae and shown to be the coccidioidal complement fixation (CF) and immunodiffusion-CF antigen. In the present study, we made use of our previously determined amino-terminal (N-terminal) sequence of the CF-chitinase to design degenerate oligonucleotide primers and to amplify and sequence a PCR product that coded for the N-terminal portion of the CF-chitinase. The PCR product was used as a hybridization probe to screen a developing spherule-(lambda)ZAP cDNA library, and three hybridizing clones were selected. These clones were converted into their pBluescript expression plasmid form in Escherichia coli and induced to express their recombinant proteins. Lysate from only one clone, pCTS 4-2A, yielded an enzymatically functional CF-chitinase and a line of identity with control immunodiffusion-CF-positive antigen. The pCTS 4-2A insert was sequenced and found to contain a deduced open reading frame coding for a 427-amino-acid polypeptide with an approximate molecular weight of 47 kDa. When purified by a chitin adsorption-desorption method, the recombinant protein exhibited virtually identical characteristics to those o...Continue Reading

References

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Citations

Sep 20, 2005·Mycopathologia·Natalia ElguezabalJosé Pontón
Jun 1, 2001·Current Infectious Disease Reports·Jiejian Lin, Richard J. Hamill
Jun 9, 2006·Mycopathologia·Li Duo-Chuan
Jun 7, 2006·Annual Review of Microbiology·Chad A Rappleye, William E Goldman
Oct 18, 2014·Mycopathologia·Suzanne M JohnsonDemosthenes Pappagianis
Dec 17, 2020·Journal of Fungi·Neil M Ampel
Dec 6, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Esperanza Duarte EscalanteMaría Del Rocío Reyes Montes
Mar 30, 2001·Fungal Genetics and Biology : FG & B·D Pappagianis
Oct 23, 1997·Clinical Microbiology Reviews·G S Deepe
Aug 3, 2021·Open Forum Infectious Diseases·Francisca J GrillDouglas F Lake
Aug 24, 2010·Autoimmunity Reviews·Andrea T Borchers, M Eric Gershwin

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