Last data update: 20 April 2021 09:48 CEST
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|
|Replicon:||Enterococcus faecalis plasmid pAD1 origin|
|Host range:||Enterococcus (formerly Streptococcus) faecalis
Gram-positive bacterial strains
|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.|
|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:||Weaver et al., J. Bacteriol. 172 (1990), 2633-2641 [PMID: 2158976]
Weaver et al., Plasmid 22 (1989), 106-119 [PMID: 2516332]
Weaver et al., J. Bacteriol. 170 (1988), 4343-4352 [PMID: 2842316]
Tomich et al., Antimicrob. Agents Chemother. 15 (1979), 828-830 [PMID: 112916]
Ehrenfeld et al., J. Bacteriol. 169 (1987), 3473-3481 [PMID: 3038841]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Enterococcus faecalis OG1X|
|Cultivation medium:||MRS + streptomycin (25 μg/ml)|
|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.