Biopolymers and Cell. 2011; 27(4): 310-313
CLONING AND EXPRESSION OF ERYSIPELOTHRIX RHUSIOPATHIAE SPAA GENE IN ESCHERICHIA COLI
Deriabin O. M., 1Deryabina O. G., 1Gilchuk P. V.
Institute of Veterinary Medicine NAAS of Ukraine
30, Donetska Str., Kyiv, Ukraine, 03151
1Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680
Aim. Cloning of Er. rhusiopathiae surface antigen – SPAA gene into plasmid vector and expression of SpaA protein in E. coli. Methods. Oligonucleotide primers design and synthesis, polymerase chain reaction, cloning, recombinant proteins expression in E. coli, electrophoresis in agarose, PAGE. Results. Full-size and truncated Er. rhusiopathiae SPAA genes have been cloned into expression plasmid pET24a(+). Expression of full-size gene hasn’t accompanied with the significant recombinant protein amount synthesis, presumably due to its toxicity for E. coli cells. Elimination of the fragment coding for proline-rich region and tandem repeats domain from full-size gene led to the obtaining of the truncated variant which produced stable synthesis of the recombinant protein with expected molecular mass. Conclusions. recombinant plasmid containing fragment of Er. rhusiopathiae SPAA gene has been cloned. It allows to obtain in E. coli recombinant protein with m. m. 47 kDa in preparative amount.
Keywords: Erysipelothrix rhusiopathiae, SPAA gene, recombinant protein