Biopolym. Cell. 2011; 27(4):310-313.
Short Communications
Cloning and expression of Erysipelothrix rhusiopathiae SPAA gene
in Escherichia coli
- Institute of Veterinary Medicine, NAAS of Ukraine
30, Donetska Str., Kyiv, Ukraine, 03151 - Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
Abstract
Aim. Cloning of Er. rhusiopathiae surface antigen – SPAA gene into plasmid vector and its expression in E. coli. Methods. Oligonucleotide primers design and synthesis, polymerase chain reaction, cloning, recombinant proteins expression in E. coli, electrophoresis in agarose, PAGE. Results. The full-size and truncated Er. rhusiopathiae SPAA genes were cloned into expression plasmid pET24a(+). The expression of full-size gene didn't result in significant recombinant protein production, presumably due to its toxicity for E. coli cells. The elimination of fragment coding for proline-rich region and tandem repeats domain from full-size gene led to production truncated variant of recombinant protein with expected molecular mass. Conclusions. The recombinant plasmid containing fragment of Er. rhusiopathiae SPAA gene was constructed. It allows obtaining preparative amount recombinant protein with m. m. 47 kDa in E. coli.
Keywords: Erysipelothrix rhusiopathiae, SPAА gene, recombinant protein
Full text: (PDF, in Russian)
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