Development of replicative oriC plasmids and their versatile use in genetic manipulation of Cytophaga hutchinsonii.

Abstract:

:Cytophaga hutchinsonii is a Gram-negative aerobic soil bacterium which can digest crystalline cellulose completely through a strategy different from that of the well-studied cellulolytic aerobic fungi and anaerobic bacteria. However, despite the availability of a published genome sequence, studies on this organism have been very limited because of the lack of a genetic manipulation system. This paper describes the development of replicative oriC plasmids, carrying the replication origin of the C. hutchinsonii chromosome, and an electroporation method for Escherichia coli-C. hutchinsonii shuttle vectors based on oriC plasmids with an efficiency of about 2 × 10(4) transformants per microgram plasmid DNA. Heterologous proteins, including green fluorescent protein and β-galactosidase, were expressed successfully and proved functional in C. hutchinsonii under the control of the CHU_1284 promoter in oriC plasmids. Finally, the gene CHU_0344, encoding the main extracellular protein, was targeted by homologous recombination based on the oriC plasmid. These genetic techniques and tools will provide a means to study the novel cellulose degradation system of C. hutchinsonii.

authors

Xu Y,Ji X,Chen N,Li P,Liu W,Lu X

doi

10.1007/s00253-011-3572-0

subject

Has Abstract

pub_date

2012-01-01 00:00:00

pages

697-705

issue

2

eissn

0175-7598

issn

1432-0614

journal_volume

93

pub_type

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