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GeneCorner host details

Last data update: 24 January 2024 16:39 CET

Host name: Escherichia coli K12 Dual7 (LMBP 13433)

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Species:Escherichia coli
Group:K12
Strain designation:Dual7
Accession number: LMBP 13433
Depositor: This host strain was deposited by Prof. Dr M. Shoulders1.
1 Department of Chemistry, Massachusetts Institute of Technology, Cambridge, USA
Containment level: This strain has been assigned the containment level Class 1 following the European Directive 2009/41/EC on the contained use of genetically modified organisms, and its updates (see also the Belgian risk group classification).
Medium: LB-Lennox + streptomycin (25 µg/ml)
Selection marker: Streptomycin
Temperature: 37°C
Genotype: F- mcrA ∆(mrr-hsdRMS-mcrBC) φ80lacZ∆M15 ∆lacX74 recA1 endA1 araD139 ∆(ara-leu)7697::[PA1lacO-Tenth MutaT7A->G] galU galK λ- rpsL nupG ∆ung::[PA1lacO-Tenth MutaT C ->T] ∆motAB ∆csgABCDEFG [PlacI<>Ptac]
Phenotype: StrR
Properties: The Dual7 strain genomically encodes two mutagenic chimeric proteins: MutaT7A→G (TadA7.10, an adenosine deaminase, fused to T7RNAP) and MutaT7C→ T (rAPOBEC1, a cytidine deaminase, fused to T7RNAP). These chimeric fusions to T7 RNA polymerase (T7RNAP), induce A-to-G and C-to-T mutations, respectively. The expression of both chimeric proteins can be induced via the addition of IPTG, since both chimeras are placed under the control of a modified IPTG-inducible promoter.
Further information: Given that the T7RNAP domain of the fusions specifically recognizes the T7 promoter and begins transcribing downstream regions, the mutagenic chimeric protiens can be used to cause mutations in a highly targeted and processive manner (i.e. on any region downstream of aT7 promoter). Thus, the Dual7 strain is broadly applicable for in vivo mutagenesis and directed evolution.

PlacI was replaced by Ptac through lambda red recombineering.

The Dual7 strain is a derivative of Escherichia coli K12 DH10B of which the full genome sequence is available at the ENA/GenBank nucleotide sequence databases under accession number CP000948.1.
Original reference: Mengiste et al., Nucleic Acids Res. 51 (2023), e31 [PMID: 36715334] [DOI: 10.1093/nar/gkad003]
Restricted use: - BCCM MTA
- The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.

Refer in your Materials and Methods:

Escherichia coli K12 Dual7 (LMBP 13433) is available at BCCM/GeneCorner. It was deposited by Prof. Dr M. Shoulders and was published in Mengiste et al., 2023.

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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