FIELD: biotechnology, genetic engineering, medicinal microbiology.
SUBSTANCE: invention describes novel recombinant plasmids comprising V. cholerae 01 biovars eltor and classicae cloned cef gene, non01/non0139 (0130 and 0139 (Bengal) - pCef61B, pCef69B, pCef49B and pCef31B as components of vector plasmid pBAD18, respectively. Plasmids are prepared by incorporation of a PCR-amplificate of Vibrio choleare cef gene by sites Eco RI - Xba I in polylinker MCS2 of vector plasmid pBAD18 in orientation providing the transcription direction under control of PBAD-promoter. Strains of E. coli - Jm103pCef61B (KM 190), Jm103pCef69B (KM 191), Jm103pCef49B (KM 189) and Jm103pCef31B (KM 188) are producers of recombinant plasmids and Cef protein prepared by transformation of E. coli strain Jm103 with recombinant plasmids. Product of cef is carried out in induction with arabinose. Recombinant proteins Cef retain all three type of activity: esterase and biological one in vitro and in vivo. Invention can be used for investigation of role of cytotonic factor cef (CHO cell elongating factor) in pathogenesis of cholera, its biochemical and biological activity.
EFFECT: improved and valuable properties of plasmid, gene and protein.
8 cl, 2 dwg, 4 ex
