The use of plasmid R1162 and derivatives for gene cloning in the methanol-utilizing Pseudomonas AM1

Gautier, Fritz; Bonewald, Renate

doi:10.1007/bf00270487

Cited by 41 publications

(14 citation statements)

References 18 publications

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“…This is probably a general property of IncQ plasmids, which fall into the IncP-4 group of P. aeruginosa (Jacoby, 1977). The combination of small size, high copy number, and broad host range has lead to efforts to develop cloning vectors from these plasmids (Nagahari & Sakaguchi, 1978;Bagdasarian et al, 1979;Barth, 1979;Gautier & Bonewald, 1980). Our interest lies in the conjugal transmission of IncQ plasmids.…”

Section: Introductionmentioning

confidence: 99%

Mobilization of the non-conjugative IncQ plasmid RSF1010

Willetts

Crowther

1981

Genet. Res.

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The broad host-range non-conjugative IncQ plasmid RSF1010 was mobilised with 100 % efficiency in membrane filter matings, both in E. coli K12 and P. aeruginosa PAO, by broad host-range conjugative IncP plasmids. No homology between RSF1010 and an IncP plasmid could be detected. In E. coli, Incla and IncX plasmids, but not IncF, IncN or IncW plasmids, were also relatively efficient at mobilising RSF1010, while in P. aeruginosa, R91-5 (IncP-10) was highly efficient, but pMG5 (IncP-2) and FP2 (IncP-8) were very inefficient. IncP plasmids also mobilised several plasmids derived from RSF1010 for use as in vectors in in vitro recombination experiments very efficiently, and pSClOl quite efficiently: this reduces the level of biological containment possible with these plasmids. 12GRH 37

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Section: Introductionmentioning

confidence: 99%

Mobilization of the non-conjugative IncQ plasmid RSF1010

Willetts

Crowther

1981

Genet. Res.

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“…Gautier and Bonewald have reported the complementation of mutant M1SA with a Methylobacterium sp. strain AM1 clone bank constructed in a broad-host-range IncQ plasmid vector (11). More recently, Allen and Hanson have been successful in complementing Methylobacterium sp.…”

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confidence: 99%

Isolation and complementation analysis of 10 methanol oxidation mutant classes and identification of the methanol dehydrogenase structural gene of Methylobacterium sp. strain AM1

Nunn¹,

Lidstrom²

1986

J Bacteriol

147

151

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A method has been developed for the direct selection of methanol oxidation mutants of the facultative methylotroph Methylobacterium sp. strain AM1 (formerly Pseudomonas sp. strain AM1). Using this direct selection technique, we have isolated mutants of Methylobacterium sp. strain AM1 that are no longer capable of growth on methanol but retain the ability to grow on methylamine. These methanol oxidation (Mox) mutants were complemented with a genomic clone bank of this organism constructed in the broad-host-range cosmid pVK100, and subcloning and TnS mutagenesis experiments have assigned the Mox mutants to 10 distinct complementation groups. Using an open reading frame beta-galactosidase fusion vector and antibodies specific for Methylobacterium sp. strain AMI methanol dehydrogenase, we have identified the methanol dehydrogenase structural gene and determined the direction of transcription. The results suggest that the synthesis and utilization of an active methanol dehydrogenase in this organism requires at least 10 different gene functions.All methylotrophic bacteria that grow on methane or methanol use methanol dehydrogenase (MDH; EC.1.1.99.8) to oxidize methanol to formaldehyde, which may then be further oxidized or assimilated into cell carbon (Fig. 1). This enzyme is a major cellular protein during growth on methane or methanol and is remarkably similar in most methylotrophs (2). However, little is known concerning the details of genetics, expression, assembly, or stability of the methanol oxidation system. We have chosen to study methanol oxidation in Methylobacterium sp. strain AM1 (formerly Pseudomonas sp. strain AM1) for a number of reasons. Strain AM1 is a facultative methylotroph capable of growth on methanol or methylamine but not methane (30), and this organism has been extensively studied and much information is available concerning the biochemistry of C1 metabolism in this organism. The MDH of Methylobacterium sp. strain AM1 has a number of features similar to those of the MDH found in most methanotrophic and methylotrophic bacteria (2, 29) and might represent a model system for the study of the genetics of methanol oxidation in a methylotroph. Perhaps most importantly, the methanol oxidation step can be genetically defined in Methylobacterium sp. strain AM1 by obtaining mutants of this organism no longer able to grow on methanol but still able to utilize methylamine as a growth substrate. Further assimilation and oxidation of formaldehyde by this organism are independent of whether methanol or methylamine is the substrate for growth (Fig. 1)

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“…Apart from transformation in methane and methanol-utilising bacteria [1,2], naturally occurring recombination in these organisms has not been reported. One method for obtaining recombinant bacteria has been to infect methylotrophs with broad host-range plasmids from Escherichia coli [3][4][5] and, although this has been used successfully to manipulate bacteria of industrial importance [4], this method has the possible disadvantage that the presence of a foreign plasmid may interfere with naturally occurring genetic processes. An alternative strategy has been to screen bacteria for plasmids which are normally present in the hope that some of these might promote conjugation.…”

Section: Introduction 2 Methodsmentioning

confidence: 99%

Isolation and characterization of a high molecular weight plasmid from the obligate methanol-utilising bacterium Methylomonas (Methanomonas) methylovora

Monteiro¹

1982

FEMS Microbiology Letters

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The use of plasmid R1162 and derivatives for gene cloning in the methanol-utilizing Pseudomonas AM1

Cited by 41 publications

References 18 publications

Mobilization of the non-conjugative IncQ plasmid RSF1010

Mobilization of the non-conjugative IncQ plasmid RSF1010

Isolation and complementation analysis of 10 methanol oxidation mutant classes and identification of the methanol dehydrogenase structural gene of Methylobacterium sp. strain AM1

Isolation and characterization of a high molecular weight plasmid from the obligate methanol-utilising bacterium Methylomonas (Methanomonas) methylovora

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