Cloning and expression of the Bacteroides fragilis YCH46 neuraminidase gene in Eschirichia coli and Bacteroides uniformis |
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Authors: | Tsuneko Ono Shigeru Akimoto Takemi Kinouchi Keiko Kataoka Yoshinari Ohnishi |
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Affiliation: | Department of Bacteriology, School of Medicine, The University of Tokushima, Kuramoto-cho, Tokushima 770, Japan |
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Abstract: | Abstract 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 Xho I- Kpn I 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. |
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Keywords: | Bacteroides fragilis Cosmid library Neuraminidase gene Shuttle vector Bacteroides uniformis |
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