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pOVEX vector: prokaryotic expression and purification of onchocerciasis vaccine candidate antigens as fusion proteins with the 24 kD Onchocerca volvulus glutathione S‐transferase
Author(s) -
Liebau Eva,
Spillner Edzard,
HenkleDührsen Kimberly
Publication year - 1997
Publication title -
tropical medicine and international health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.056
H-Index - 114
eISSN - 1365-3156
pISSN - 1360-2276
DOI - 10.1046/j.1365-3156.1997.d01-352.x
Subject(s) - onchocerca volvulus , fusion protein , biology , glutathione s transferase , antigen , onchocerciasis , vector (molecular biology) , sepharose , glutathione , microbiology and biotechnology , immunology , recombinant dna , biochemistry , enzyme , gene
An expression vector, pOVEX, has been designed and constructed, combining the advantages of the expression vectors pGEX‐3X and pJC20. The pOVEX vector produces a fusion protein with the 24 kD Onchocerca volvulus glutathione S‐transferase (OvGST2) which is easy to purify in one step from bacterial extracts under non‐denaturing conditions using glutathione‐sepharose chromatography. High yields of fusion protein were produced from this T7 RNA polymerase‐dependent expression vector, which were then cleaved by digestion with the factor Xa protease to separate the OVGST2 polypeptide from the expressed protein of interest. This vector will be particularly useful to O. volvulus investigators for the production of O. volvulus antigens for the analyses of host humoural and cellular responses to these proteins and for immunization studies.