[Expression and purification of the recombinant fusion protein GST-cysteine protease of Ancylostoma ceylanicum].
cDNA encoding Ancylostoma ceylanicum cysteine protease (ACEY-1) was cloned into pGEX-4T eucaryotic expression vector then E. coli BL2 cells were transformed with the plasmid using electroporation method. The expression of the recombinant protein in liquid cultures was induced by the addition of IPTG. The Western blot analysis showed that the recombinated ACEY-1 may be solublised only partially at pH 7.4 which allowed affinity-purification of this protein using glutathione-sepharose column.
Wiśniewski
Marcin
M
Zakład Parazytologii i Inwazjologii, SGGW, Warszawa.
Mieszczanek
Juliusz
J
Widrychowicz
Halina
H
D000722:Ancylostoma / Q000201:enzymology*; D000818:Animals; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000302:isolation & purification*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification*
D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D003546:Cysteine Endopeptidases