RECOMBINANT PLASMID pET40CmAP/ MPF DNA, ENCODING BIFUNCTIONAL HYBRID CmAP/OmpF POLYPEPTIDE WITH PROPERTIES OF HIGHLY-ACTIVE ALKALINE PHOSPHATASE CmAP AND PORE-FORMING MEMBRANE OmpF PROTEIN, AND RECOMBINANT STRAIN OF E. COLI ROSETTA (DE3)/pET40CmAP/OmpF - PRODUCER OF BIFUNCTIONAL HYBRID CmAP/OmpF POLYPEPTIDE

摘要

FIELD: biotechnology.;SUBSTANCE: said plasmid is characterized by presence of the following fragments: a NcoI/SalI fragment of pET-40b(+) plasmid (Novagen) and a 2577 bp DNA fragment containing a chimeric gene encoding the full-length alkaline phosphatase CmAP and the full-length Yersinia pseudotuberculosis OmpF porin, linked together by a sequence encoding a flexible linker (G₄S)₃ (SEQ ID NO: 1). A recombinant strain of E. coli Rosetta (DE3)/pET40CmAP/OmpF, which is a producer of a bifunctional hybrid CmAP/OmpF polypeptide, and which is obtained by modification of the E. coli Rosetta (DE3) strain with this pET40CmAP/OmpF plasmid, is proposed.;EFFECT: high yield of hybrid protein with the properties of recombinant highly-active alkaline phosphatase and pore-forming protein.;2 cl, 2 dwg, 3 ex