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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Heterologous expression of genes in the anaerobic bacterium Streptococcus bovis

Ekinci, Mehmet Sait January 1997 (has links)
The main objective of this work was to investigate the expression of xylanase and cellulose genes from <I>Ruminococcus flavefaciens</I> when introduced into related Gram-positive bacteria including the rumen bacterium <I>Streptococcus bovis</I>. In addition the discovery, and isolation of the β(1,3-1,4)-glucanase gene of <I>S. bovis</I> in the course of this work may provide new possibilities to express foreign genes in <I>S. bovis</I> or in other Gram-positive bacteria. The main findings of this work are summarised below: 1. Genes encoding polysaccharidase activity from the strictly anaerobic rumen bacterium <I>R. flavefaciens</I> can be transferred by electroporation into the ruminal bacterium <I>S. bovis</I> and into other Gram-positive bacteria from different habitats, including <I>Lactococcus lactis, Enterococcus faecalis </I>and <I>Streptococcus sanguis</I>. 2. <I>XynD </I>and <I>endA</I> genes of <I>R. flavefaciens </I>can be expressed from their own promoters in these species. Among the bacteria used as hosts for gene expression, <I>S. bovis </I>gave the highest yields of active enzyme. The expression levels of both gene products were found to be higher in <I>S. bovis</I> than in <I>E. coli</I>. 3. The <I>R. flavefaciens </I>enzymes were mainly secreted in the culture medium of <I>S. bovis</I>; however in <I>E. coli</I> they were mainly cell-associated. 4. The full-length enzyme of <I>xynD</I> was detected in several Gram-positive bacteria, suggesting the effect of proteases may be less than in <I>E. coli</I>. 5. The general rearrangement of the introduced plasmid and genes were not found in Gram-positive bacteria and the genes seem to be stable in these organisms. However rearrangement of <I>xynD</I> was observed in some transformants of <I>S. bovis </I>JB1, although non-rearranged transformants were also obtained. 6. Expression of <I>endA</I> and <I>xynD</I> activity was affected by energy sources supplied to <I>S. bovis</I> cultures, reflecting the accumulation of lactic acid.

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