pOVEX vector: prokaryotic expression and purification of onchocerciasis vaccine candidate antigens as fusion proteins with the 24 kD Onchocerca volvulus glutathione S-transferase.
An expression vector, pOVEX, has been designed and constructed, combining the advantages of the expression vectors pGEX-3X and pJC2o. 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 humoral and cellular responses to these proteins and for immunization studies.
Liebau
E
E
Department of Biochemistry, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany.
Spillner
E
E
Henkle-Dührsen
K
K
D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics* / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics* / Q000276:immunology*; D005982:Glutathione Transferase / Q000235:genetics* / Q000276:immunology*; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017181:Onchocerca volvulus / Q000201:enzymology / Q000235:genetics* / Q000276:immunology; D010957:Plasmids / Q000235:genetics / Q000276:immunology; D011387:Prokaryotic Cells; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D014614:Vaccines, Synthetic / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D000947:Antigens, Helminth; D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D005982:Glutathione Transferase