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E. coli K12 ER1821 |
 | | | | E. coli K12 ER1821 | | | Limit of 1 per order | |
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Prices are in US dollars and valid only for US orders.
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Description:
MM294 background: McrA- McrBC- EcoK r- m- Mrr- . A suspension of E. coli ER1821 which has been grown in rich medium and brought to 50% glycerol.
Genotype: F- glnV44 e14-(McrA-) rfbD1? relA1? endA1 spoT1? thi-1 Δ(mcrC-mrr)114::IS10
Recommended Growth Medium: LB
Growth Temperature: 37°C
Strain Properties
dam/dcm Methylation:
dam+
, dcm+
Protease-deficient:
No
Laclq:
No
Storage Conditions
Storage Temperature:
-20°C
For long term storage (>30 days), store at -70°C.
Notes
General notes:- Restriction Defects: McrA, McrBC, and Mrr all restrict methylcytosine-containing DNA methylated by the CpG methylase (M.Sss I) (1). In addition, McrA and McrBC specifically restrict DNA modified by different sets of more sequence-specific cytosine methylases (2).
Mrr also restricts many methyladenine-containing sequences (3,4). Restriction by each of these three restriction systems individually (tested with λ DNA modified with the Sss I methylase and then packaged into phage) is: McrA, 250-fold; McrBC, 100-fold; Mrr, 200-fold. Cumulative restriction when all three are present in the same cell is 5,000-fold (1). In addition, EcoK restricts DNA not modified at K sites by 10-fold to 5,000-fold, depending on the number of sites (5). ER1821 lacks all four activities.
Usage notes:- Rubidium Chloride Method: RbCl Transformation Protocol
References
- Kelleher, J.E. and Raleigh, E.A. (1991) J. Bacteriol., 173, 5220-5223.
- Raleigh, E.A. and Wilson, G. (1986) Proc. Natl. Acad. Sci. USA, 83, 9070-9074.
- Waite-Rees, P.A. et al. (1991) J. Bacteriol., 173, 5207-5219.
- Heitman, J. and Model, P. (1987) J. Bacteriol., 169, 3243-3250.
- Murray, N.E. et al. (1973) Mol. Gen. Genet., 120, 261-281.
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