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Cloning, expression and purification of recombinant streptokinase: partial characterization of the protein expressed in Escherichia coli BJMBR
Avilán,L.; Yarzábal,A.; Jürgensen,C.; Bastidas,M.; Cruz,J.; Puig,J..
We cloned the streptokinase (STK) gene of Streptococcus equisimilis in an expression vector of Escherichia coli to overexpress the profibrinolytic protein under the control of a tac promoter. Almost all the recombinant STK was exported to the periplasmic space and recovered after gentle lysozyme digestion of induced cells. The periplasmic fraction was chromatographed on DEAE Sepharose followed by chromatography on phenyl-agarose. Active proteins eluted between 4.5 and 0% ammonium sulfate, when a linear gradient was applied. Three major STK derivatives of 47.5 kDa, 45 kDa and 32 kDa were detected by Western blot analysis with a polyclonal antibody. The 32-kDa protein formed a complex with human plasminogen but did not exhibit Glu-plasminogen activator...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Recombinant streptokinase; Plasminogen activators; Protein purification.
Ano: 1997 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1997001200007
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