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GeneCorner host details

Last data update: 24 January 2024 16:39 CET

Host name: Escherichia coli K12 Dual8 (LMBP 13434)

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Species:Escherichia coli
Group:K12
Strain designation:Dual8
Accession number: LMBP 13434
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 eMutaT C ->T] ∆motAB ∆csgABCDEFG [PlacI<>Ptac]
Phenotype: StrR
Properties: The Dual8 strain genomically encodes two mutagenic chimeric proteins: eMutaT7A→G (TadA8e, 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 proteins can be used to cause mutations in a highly targeted and processive manner (i.e. on any region downstream of a T7 promoter). Thus, the Dual8 strain is broadly applicable for in vivo mutagenesis and directed evolution.

PlacI was replaced by Ptac through lambda red recombineering.

The Dual8 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 Dual8 (LMBP 13434) 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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