DNA FRAGMENT CORRESPONDING TO GENE SKC-2, METHOD FOR EXPRESSING DNA FRAGMENT CORRESPONDING TO GENE SKC-2, PLASMID VECTOR PEKG3 PROVIDING FOR EXPRESSING DNA FRAGMENT CORRESPONDING TO GENE SKC-2, PLASMID VECTOR PES KC-4 PROVIDING FOR EXPRESSING DNA FRAGMENT CORRESPONDING TO GENE SKC-2, ESCHERICHIA COLI BACTERIA STRAIN HAVING PLASMID VECTOR PEKG3 USABLE FOR EXPRESSING DNA FRAGMENT CORRESPONDING TO GENE SKC-2, AND STREPTOKINASE PRODUCED IN UTILIZING ESCHERICHIA COLI BACTERIA STRAIN HSK-M

摘要

FIELD: microbiology; biological engineering; gene engineering. SUBSTANCE: method involves amplifying given fragment by means of synthetic oligonucleotides SK1, SK2 and SK3 and building-in plasmid vector for transforming competent forms of Escherichia coli bacteria strains or yeast Pichia pastoris. Vectors pEKG3 and PES KC-4 provide expression of the mentioned fragment in Escherichia coli and Pichia pastoris cells, respectively. Competent strain of Escherichia coli bacteria having plasmid vector pECG3 are used for producing streptokinase. Strain cells form colonies of irregular form and creme-like consistency with boundaries as small blades on solid medium. Maximum level of streptokinase expression is achieved in 14 h of culturing. To purify streptokinase, the cells are collected by centrifuging and destroyed with ultrasound. The produced streptokinase has molecular mass of 47000 Da, its amino acid sequence fully coincides with that grown on the base of SKC-2 gene. EFFECT: enhanced reliability of method. 8 cl, 3 dwgd