Transformation and Stability of Cloned Polysaccharidase Genes in Gram-Positive Bacteria


Abstract: Polysaccharidase genes from rumen bacteria were transferred to and expressed in ruminal and non-ruminal Gram-positive bacteria. The transformation efficiency and genetic stability of polysaccharidase genes in bacteria from different habitats were investigated. PCR amplification of cloned polysaccharidase genes from Escherichia coli, Lactococcus lactis, Enterococcus faecalis, Streptococcus sanguis and S. bovis strain 26 showed that rearrangement of plasmid and the gene fragment did not occur. The DNA band sizes from all primers agreed with the expectations of some transformants of S. bovis JB1 being rearranged. In spite of the rearrangement in S. bovis JB1 polysaccharidase activities were conserved. Growing in the continuous culture proved the plasmid survival and gene stability of the recombinant microorganisms.

Keywords: Polysaccharidase Gene, Stability, Transformation, Enzyme

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