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GeneCorner plasmid details

Last data update: 27 November 2020 04:15 CET

Plasmid name: pAD1 (LMBP 3002)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: not available
Sequence
analysis results
Genecorner:

-

Cloned DNA: -
Promoter: -
Ribosome
binding site:
-
Terminator: -
Selection marker: -
Replicon: Enterococcus faecalis plasmid pAD1 origin
Host range: Enterococcus (formerly Streptococcus) faecalis
Gram-positive bacterial strains
Parental clone: -
Further information: Plasmid pAD1 is a pheromone-responsive, conjugative plasmid from Streptococcus faecalis subsp. zymogenes strain DS16.
It contains the hemolysin-bacteriocin determinant (cytolysin) and provides resistance to UV.
It doesn't carry a selectable marker.
pAD1 is a very large plasmid, i.e. 59,6 kb; consequently, the DNA is very fragile and easily damaged.
Being a conjugative plasmid, it is most probably a low-copy number plasmid.
Referring to Ehrenfeld et al (1987), it should be possible to isolate pAD1 DNA according to a lysozyme-alkaline based method. In this context, a large-volume DNA prep might help to isolate low-copy number plasmids. Better results might be obtained by equilibrium ultracentrifugation of Triton lysates or Sarkosyl lysates in a cesium chloride-ethidium bromide gradient (as mentioned in Clewell et al. (1982)), but please take into account that this technique is very costly as well as cumbersome and requires some hands on experience. Commercial kits for the isolation of large plasmids might also be useful.
EMBL Accession number: -
Latest sequence update: 17/09/2001
Authenticity test: By lack of sequence data, it was not possible to perform the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr D.B. Clewell(1).
(1) University of Michigan, USA
Plasmid reference: Clewell et al., J. Bacteriol. 152 (1982), 1220-1230 [PMID: 6292164]
Related plasmid reference: Ehrenfeld et al., J. Bacteriol. 169 (1987), 3473-3481 [PMID: 3038841]
Tomich et al., Antimicrob. Agents Chemother. 15 (1979), 828-830 [PMID: 112916]
Weaver et al., J. Bacteriol. 170 (1988), 4343-4352 [PMID: 2842316]
Weaver et al., Plasmid 22 (1989), 106-119 [PMID: 2516332]
Weaver et al., J. Bacteriol. 172 (1990), 2633-2641 [PMID: 2158976]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Enterococcus faecalis OG1X
Host reference: -
Cultivation medium: MRS + streptomycin (25 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L2
Cultivation remark: Cultivate for at least 20 hours. The strain also grows on selective Slanetz-Bartley and M17 agar.
Other culture collection numbers: -

Refer in your Materials and Methods:

pAD1 (LMBP 3002) is available at BCCM/GeneCorner. This plasmid was deposited by Dr D.B. Clewell and was published in Clewell et al., 1982.

Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.

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