Cell

Part:BBa_K615002

Designed by: Naomi Genuth   Group: iGEM11_Harvard   (2011-09-26)

EcNR2 E. coli strain

The EcNR2 strain is a modified E. coli strain which has been designed for easy genome manipulation. It contains the lambda red machinery and the MutS knockout needed for efficient lambda red and multiplex automated genome engineering (MAGE).

See below for a summary of strain features and techniques:

HARVEcnr2summary.png

Usage and Biology

The EcNR2 strain is ideal for the techniques of lambda red and multiplex automated genome engineering (MAGE). Lambda red permits the insertion of large pieces of DNA into the genome, while MAGE can create small point mutations, insertions, and deletions. Both lambda red and MAGE were used successfully on this strain by the 2011 Harvard iGEM team to create two one-hybrid systems, submitted as chassis BBa_K615000 and BBa_K615003. See the figures below for examples of successful lambda red and MAGE procedures performed using this strain.

HARVlambda.png


HARVMAGEresults2.png

For detailed lambda red and MAGE protocols, please see the 2011 Harvard iGEM team's [http://2011.igem.org/Team:Harvard/Protocols Protocols] page. For descriptions of how these genome editing techniques work, go to the [http://2011.igem.org/Team:Harvard/Technology/MAGE MAGE] and [http://2011.igem.org/Team:Harvard/Lambda_Red Lambda Red] pages.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
biologygrow only at 30⁰C
genotypeMG1655 with λ-prophage::bioA/bioB and cmR::mutS
resistancechloramphenicol, ampicillin