Cloning and expression of the Bacteroides fragilis YCH46 neuraminidase gene in Escherichia coli and Bacteroides uniformis.
A neuraminidase-encoding gene nanH of Bacteroides fragilis strain YCH46 was cloned into the cosmid vector pHC79. The nanH gene was subcloned from the cosmid and was located within a 2.2-kb XhoI-KpnI fragment. Southern hybridization experiments demonstrated that the gene was present as a single copy on the bacterial chromosome. Neuraminidase activity expressed in the initial Escherichia coli clone was approximately 3600-fold lower than that expressed in B. fragilis YCH46. However, when nanH was transferred from E. coli to B. uniformis by mobilization of a shuttle plasmid, the transconjugant expressed 1100-fold higher activity than the E. coli donor did. These results suggest that modes of nanH expression in E. coli and Bacteroides are heterologous.
Ono
T
T
Department of Bacteriology, School of Medicine, University of Tokushima, Japan.
Akimoto
S
S
Kinouchi
T
T
Kataoka
K
K
Ohnishi
Y
Y
D001439:Bacteroides / Q000235:genetics; D001441:Bacteroides fragilis / Q000201:enzymology / Q000235:genetics*; D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D009439:Neuraminidase / Q000235:genetics*
D004269:DNA, Bacterial; D009439:Neuraminidase