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Useof pEX and pGEX bacterial heterologous protein expression systems forrecombinant oncoprotein production and antisera generation
Author(s) -
Pompéia Celine,
Ortis Fernanda,
Armelin Mari C. S.
Publication year - 1997
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1997.tb00434.x
Subject(s) - fusion protein , recombinant dna , antiserum , heterologous , biology , microbiology and biotechnology , heterologous expression , biochemistry , gene , chemistry , antibody , genetics
In order to study the role played by known and novel genes in growth control and neoplasia, we here compare the pEX and pGEX bacterial expression systems for recombinant oncoprotein production and for generation of specific antisera. The results of five pEX (MS2‐c‐Fos, MS2‐Fra‐1, MS2‐JunD, bgal‐c‐Jun and bgal‐JunB) and two pGEX [glutathione S‐transferase (GSH)‐JE/MCP‐1 and GST‐JunD] fusion‐protein productions are presented. Higher (15Ő43‐fold) yields are obtained with the pEX system, but only the pGEX system allows separation of the protein of interest from the fusion moiety by digestion with specific proteases. The degree of fusion‐protein purification, as assessed by SDS/PAGE, is similar for both systems. Proteins produced by both systems were successfully used in the generation of specific antisera. The choice between the pEX and pGEX systems is dependent upon the specific recombinant protein produced.