Generator

Part:BBa_K117001:Design

Designed by: Nguyen Xuan Hung   Group: iGEM08_NTU-Singapore   (2008-10-07)
Revision as of 16:13, 29 October 2008 by Dariusaw (Talk | contribs) (→‎References)

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ColicinE7 with Immunity


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]


Design Notes

E7 Colicin (CEA) and Lysis (CeI) gene are extracted from E Coli bacteria strain BW3110 and BW containing the plasmid ColE7-K317. Plasmid ColE7-k317 is extracted from the bacteria stain using Qiagen plasmid extraction kit. The desired sequence of CEA and CeL is then extracted from the plasmids using PCR and oligo fragments of 5'-ATGAGCGGTGGAGATGGA-3' and 3'-CCTAAATTTGTCCCGACT-5' with the biobricks prefix and suffix flanking the oligo fragments respectively. Through PCR method, biobrick prefix and suffix are added to either sides of the CEA and CeL gene, standardizing the gene sequence for future biobricks combination.

Source

Bacteria strain W3110 and BW that contain the plasmid ColE7-K317 was generously donated by Professor Chak Kin Fu from National Yang Ming University, Taipei. The above mentioned E coli strains are derivatives of the E Coli K12 strain. Oligo Fragments were ordered through and synthesized by AITbiotech.

References

[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=144374 Gene sequence from NCBI]

Cloning and Characterization of ColE7 Plasmid. J Gen Microbiol 137:97-100

Characterization of the cea gene of the ColE7 Plasmid. Mol Gen Genet (1992) 233:177-183 "