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Volume 147, Issue 3

Development of a genetic system for the transfer of DNA into

The GenBank accession number for the sequence reported in this paper is AF221716.

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Abstract

(now ) is a Gram-negative soil bacterium which can produce yellow pigments. It synthesizes five enzymes that degrade glycosoaminoglycan molecules. The study of this unique bacterium has been limited by the absence of a genetic manipulation system. In this paper, the construction of a conjugation/integration plasmid system and a broad-host-range plasmid, both of which contain a functional selective marker created by placing the trimethoprim resistance gene, , under the control of the regulatory region is described. Both plasmids were introduced into by conjugation and/or electroporation, and trimethoprim resistant colonies were obtained. Fifty electroporants were obtained per microgram covalently closed circular plasmid DNA. The existence of integrated plasmid DNA was confirmed by Southern hybridization and PCR. The existence of a derivative of the broad-host-range plasmid pBBR1 in was demonstrated by plasmid digestion and Southern hybridization, and by transformation of .

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Keyword(s): Ap, ampicillin , Cm, chloramphenicol , Em, erythromycin , Flavobacterium heparinum , Gm, gentamicin , heterologous DNA transfer , HT, trimethoprim-resistance gene (cassette) , integration vector , Km, kanamycin , plasmid replication , Tc, tetracycline and Tp, trimethoprim
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2001-03-01
2024-04-18
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Development of a genetic system for the transfer of DNA into Flavobacterium heparinumThe GenBank accession number for the sequence reported in this paper is AF221716.
Microbiology 147, 581 (2001); https://doi.org/10.1099/00221287-147-3-581
/content/journal/micro/10.1099/00221287-147-3-581
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