METHOD TO PRODUCE IMMUNOGENIC RECOMBINANT EXTRACELLULAR FRAGMENT CONNEXIN-43 Russian patent published in 2011 - IPC C12N15/00 C12N15/03 C07K1/16 C07K14/46 

FIELD: medicine.

SUBSTANCE: invention may find application in methodology for creation of biologically active antibodies joined with live cell, address delivery of medicinal agents to target cells. Proposed method includes cloning of the second extracellular loop E2 Cx43, representing a fragment of connexin-43 Q173 -1208 with aminoacid sequence QWYIYGFSLSAVYTCKRDPCPHQVDCFLSRPTEKTI, into two plasmid vectors pCBDQ and pHPMLQ. Then transformation of E.coli cells is carried out with produced plasmid DNAs. Then transformed cells are cultivated up to optical density of 0.7-0.9. Then plasmid expression 0.9-1.1 mM is induced with isopropyl thiogalactoside. Afterwards recombinant fused proteins and their release in presence of 7.9-8.1 M of urea by method of chromatography at Ni-NTA agarose.

EFFECT: proposed invention makes it possible to develop method to produce immunogenic recombinant extracellular fragment of rat connexin-43 in composition of highly immunogenic chimeric protein suitable to develop monoclonal antibodies.

2 dwg, 1 tbl, 1 ex