Isolation of Inc P-2 plasmid DNA from Pseudomonas aeruginosa

Fennewald, Michael; Prevatt, William; Meyer, Richard J.; Shapiro, James A.

doi:10.1016/0147-619x(78)90036-7

Cited by 41 publications

(18 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The highly fluorescent dye, Hoechst 33258, which binds preferentially to AT-rich DNA sequences, has been employed successfully in CsCl gradients to accentuate the buoyant density differences of cellular DNA (12,31,38). Excellent resolution of the three 0. luteus cellular DNA species present in crude cell lysates is obtained in a one-step purification/separation procedure.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of Chloroplast DNA from the Marine Chromophyte, Olisthodiscus luteus: Electron Microscopic Visualization of Isomeric Molecular Forms

Aldrich¹,

Cattolico²

1981

Plant Physiol.

View full text Add to dashboard Cite

Chloroplast DNA (ctDNA) from the marine chromophytic alga, Olisthodiscus luteus, has been isolated using a whole cel lysis method folowed by CsCI-Hoechst 33258 dye gradient centrifugation. This DNA, which has a buoyant density of 1.691 grams per cubic centimeter was identified as plastidic in origin by enrichment experiments. Inclusion of the nuclease inhibitor aurintricarboxylic acid in all lysis buffers was mandatory for isolation of high molecular weight DNA. Long linear molecules (40 to 48 micrometers) with considerable internal organization comprised the majority of the ctDNA isolated, whereas supertwisted ctDNA and open circular molecules averaging 46 micrometers were occasionally present. Also observed in this study were folded ctDNA molecules with electron dense centers ("rosettes") and plastid DNA molecules which have a tightly wound "key-ring" center. The ctDNA of Olisthodiscus has a contour length that is median to the size range reported for chiorophytic plants.A minor component of the total celular DNA, which originates from a DNase insensitive cellular structure, has a buoyant density of 1.694 grams per cubic centimeter. This DNA consists predominantly oflinear molecules, but open circles 11.5 micrometers in length and rare 22-micrometer dimers were also present.This study represents the first analysis of the extranuclear DNA of a chromophytic alga.

show abstract

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of Chloroplast DNA from the Marine Chromophyte, Olisthodiscus luteus: Electron Microscopic Visualization of Isomeric Molecular Forms

Aldrich¹,

Cattolico²

1981

Plant Physiol.

View full text Add to dashboard Cite

show abstract

“…Mapping experiments had indicated that the two loci (Benson et al, 1977 ; are transcribed towards each other and are separated by about 40 kb (Fennewald et al, 1979 ;Owen, 1986). The GjC content of the alk genes is much lower than that of the host strain and the OCT plasmid (Fennewald et al, 1978 ;Kok et al, 1989b ;Panke et al, 1999b), suggesting that these genes are part of a 55 kb mobile element, which integrated in the OCT plasmid. This appears to be a common finding for catabolic genes, and many catabolic transposons have now been described (Tan, 1999).…”

Section: Introductionmentioning

confidence: 99%

Analysis of Pseudomonas putida alkane-degradation gene clusters and flanking insertion sequences: evolution and regulation of the alk genes The EMBL accession numbers for the sequences reported in this paper are AJ245436 [P. putida (oleovorans) GPo1 alk gene clusters and flanking DNA], AJ233397 (P. putida P1 alk gene clusters and flanking DNA), AJ249793 (P. putida P1 nahKJ genes), AJ249825 [P. putida (oleovorans) GPo1 16S RNA gene] and AJ271219 (P. putida P1 16S RNA gene).

et al. 2001

View full text Add to dashboard Cite

The Pseudomonas putida GPo1 (commonly known as Pseudomonas oleovorans GPo1) alkBFGHJKL and alkST gene clusters, which encode proteins involved in the conversion of n-alkanes to fatty acids, are located end to end on the OCT plasmid, separated by 97 kb of DNA. This DNA segment encodes, amongst others, a methyl-accepting transducer protein (AlkN) that may be involved in chemotaxis to alkanes. In P. putida P1, the alkBFGHJKL and alkST gene clusters are flanked by almost identical copies of the insertion sequence ISPpu4, constituting a class 1 transposon. Other insertion sequences flank and interrupt the alk genes in both strains. Apart from the coding regions of the GPo1 and P1 alk genes (80-92 % sequence identity), only the alkB and alkS promoter regions are conserved. Competition experiments suggest that highly conserved inverted repeats in the alkB and alkS promoter regions bind AlkS.

show abstract

“…Plasmid DNA was isolated by the cleared lysate technique and cesium chloride-ethidium bromide centrifugation as described previously (8). Total DNA was made from Glu-grown PBTO cultures as described previously (15,18).…”

Section: Methodsmentioning

confidence: 99%

Variation in the ability of Pseudomonas sp. strain B13 cultures to utilize meta-chlorobenzoate is associated with tandem amplification and deamplification of DNA

Rangnekar

1988

J Bacteriol

View full text Add to dashboard Cite

strain B13 (6, 7) and Pseudomonas putida AC858 (2, 4) which degrade Ben are, however, also capable of degrading meta-chlorobenzoate (3CB). In P. putida AC858, the 3CB+ determinants are located on a 117-kilobase (kb) plasmid, pAC25 (2). A 4.3-kb DNA region on this plasmid encodes three key chlorocatechol pathway enzymes (pyrocatechase II, cycloisomerase II, and hydrolase II) for 3CB utilization (10). On the other hand, the genetic and molecular functions involved in the utilization of Ben and 3CB by the B13 strain are still unclear. The 3CB+ determinants of B13 have been shown to be transferrable by conjugation to P. putida strains (16,20,21), but it is still unclear whether these determinants are carried on a plasmid in B13. DNA hybridization studies have revealed a high degree of homology between the 3CB+ DNA in B13 and AC858 (2). Furthermore, comparison of the nucleotide sequence of the 4.3-kb 3CB+ DNA from pAC25 with the amino acid sequence of the B13 enzymes responsible for chlorocatechol utilization suggests that the 3CB+ determinants in these two different strains are identical (10). Recently, the 3CB+ DNA from B13 has been cloned and expressed in plasmid vectors (27); this clone from B13 is, however, structurally different, especially with respect to the restriction endonuclease sites, from the 3CB+ DNA cloned from AC858.The elucidation of different functions involved in the elaboration of a pathway requires the use of mutants, each of which is defective in one or more of these functions. phenotype of B13 either is not uniformly expressed or is not stably inherited in every single-colony isolation. The question is whether the phenotypic variation is a consequence of genetic alterations, and if so, the variant phenotype would be expected to be inherited and maintained in all single-colony isolates of the variant. In an effort to resolve these questions, I studied the lineages of B13 colonies, and this report describes my observations on the variation in the ability of B13 colonies to utilize Ben and 3CB as sole carbon sources. The variation in phenotypic characteristics was also documented in biochemical studies, and variation in 3CB utilization was associated with tandem amplification or deamplification of the 4.3-kb 3CB+ DNA.

show abstract

Isolation of Inc P-2 plasmid DNA from Pseudomonas aeruginosa

Cited by 41 publications

References 18 publications

Isolation and Characterization of Chloroplast DNA from the Marine Chromophyte, Olisthodiscus luteus: Electron Microscopic Visualization of Isomeric Molecular Forms

Isolation and Characterization of Chloroplast DNA from the Marine Chromophyte, Olisthodiscus luteus: Electron Microscopic Visualization of Isomeric Molecular Forms

Variation in the ability of Pseudomonas sp. strain B13 cultures to utilize meta-chlorobenzoate is associated with tandem amplification and deamplification of DNA

Contact Info

Product

Resources

About