Genetically modified yeast cell and improved process for production of clot-specific streptokinase
AssigneeCouncil of Scientific and Industrial Research
InventorsGirish Sahni, Kishore Kumar Joshi
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Abstract
Disclosed herein is an expression system for the production and secretion of biologically active clot-specific streptokinase (CSSK) protein in methylotrophic yeast. Yeast-expressed CSSK protein displays improved plasminogen activation and fibrin selectivity. Further disclosed are methylotrophic yeast transformed with at least one copy of functional cDNA sequence encoding CSSK adjunct with modified signal sequence which results in secretion of mature and correctly processed CSSK.
Claims19
An eukaryotic expression cassette comprising a polynucleotide, said polynucleotide comprising a yeast methanol inducible alcohol oxidase I promotor sequence or a yeast methanol inducible alcohol oxidase II promoter sequence, a modified Pichia alpha signal gene sequence, wherein the modified Pichia alpha signal gene sequence is an alpha signal gene sequence devoid of the STE13 protease cleavage site, a nucleic acid sequence encoding clot specific streptokinase and a transcription terminator sequence, wherein the nucleic acid sequence encoding clot specific streptokinase is selected from the group consisting of a nucleotide sequence with 100% identity to the sequence set forth in SEQ ID NO:
- 2, a nucleotide sequence with at least 85% identity to the sequence set forth in SEQ ID NO:
- 4, a nucleotide sequence with at least 85% identity to the sequence set forth in SEQ ID NO:
- 6 and a nucleotide sequence with at least 85% identity to the sequence set forth in SEQ ID NO:
- 8.
A polynucleotide having a nucleic acid sequence selected from the group consisting of SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, SEQ ID NO:
- 23, SEQ ID NO:
- 26 and SEQ ID NO:
- 28.
Dependencies are inferred from the claim text.