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KMID : 0578319910010020187
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Molecules and Cells
1991 Volume.1 No. 2 p.187 ~ p.192
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High-level Expression of Degraded Prodict-free Streptokinase in Escherichia coli by Removal of Its Putative Leader Sequence
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Park, Seung Kook
Jang, Jeong Su/Kim, Jee Cheon/Chun, Moon Jin/Byun, Si Myung
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Abstract
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The expression vector for streptokinase has been constructed from our previously cloned streptokinase coding gene (skc). Because of its deleterious effect oil the host cell growth, the leader sequence of skc was removed and the leader sequence-deleted skc was subctoned into the vector pKK223-3, which contains the regulatable tac promoter and rrnB T,T2 transcription terminator, with a short synthetic oligonucleotide adapter. When this vector, pKS601 having skc gene, was expressed in E.scherichia coli, a 47.4-kDa protein was found to be newly accumulated to about 12% of the total cellular proteins and it was identified as the streptokinase by immunoblotting with rabbit anti-streptokinase polyclonal serum. The expressed streptokinase was free from carboxyl-terminal degraded 44-kDa streptokinase and purified to near homogeneity using DEAE-cellulose and Sephadex G-150 columns. Its specific activity was about 1.3 X 105 CLN units/mg protein.
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KEYWORD
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