IGEM:PennState/Labbook/GalenLynch/2007-6-26

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Contents

July 26th, 2007: 10:00-5:45

Lambda bacteriophage regions of interest:

Lambda control region (OR)

  • Contains OR1, OR2, and OR3, shown in reverse order (i.e. the leftmost binding region below is OR3)
  • Bi directional promoter
    • in the lambda phage, CI is to the left(PRM), cro is to the right(PR)
  • recognized by cro, CI
    • cro binding affinities: OR3, OR2 ~ OR1 (OR1 is slightly less)
    • CI binding affinites: OR1, OR2 ~ OR3 (OR3 is slightly less)
  • CI acts as a positive regulator for PRM when bound to OR2, but and a negative regulator for PR when bound to OR3

ACGTTAAATC TATCACCGCA AGGGATAAAT ATCTAACACC GTGCGTGTTG ACTATTTTAC
CTCTGGCGGT GATAATGGTT GC
Cro and CI binding regions are depicted in green, -10 and -35 sequences in cyan, and shared regions are in blue

Lambda Repressor (CI)

  • Forms dimers
    • Dimers associate to form tetramers, octamers etc
  • Binds to OL and OR operators in bacteriophage genome
  • Cleaved by activated RecA protein (activates with ssDNA exposure)

ATGAGCACAA AAAAGAAACC ATTAACACAA GAGCAGCTTG AGGACGCACG TCGCCTTAAA
GCAATTTATG AAAAAAAGAA AAATGAACTT GGCTTATCCC AGGAATCTGT CGCAGACAAG
ATGGGGATGG GGCAGTCAGG CGTTGGTGCT TTATTTAATG GCATCAATGC ATTAAATGCT
TATAACGCCG CATTGCTTGC AAAAATTCTC AAAGTTAGCG TTGAAGAATT TAGCCCTTCA
ATCGCCAGAG AAATCTACGA GATGTATGAA GCGGTTAGTA TGCAGCCGTC ACTTAGAAGT
GAGTATGAGT ACCCTGTTTT TTCTCATGTT CAGGCAGGGA TGTTCTCACC TGAGCTTAGA
ACCTTTACCA AAGGTGATGC GGAGAGATGG GTAAGCACAA CCAAAAAAGC CAGTGATTCT
GCATTCTGGC TTGAGGTTGA AGGTAATTCC ATGACCGCAC CAACAGGCTC CAAGCCAAGC
TTTCCTGACG GAATGTTAAT TCTCGTTGAC CCTGAGCAGG CTGTTGAGCC AGGTGATTTC
TGCATAGCCA GACTTGGGGG TGATGAGTTT ACCTTCAAGA AACTGATCAG GGATAGCGGT
CAGGTGTTTT TACAACCACT AAACCCACAG TACCCAATGA TCCCATGCAA TGAGAGTTGT
TCCGTTGTGG GGAAAGTTAT CGCTAGTCAG TGGCCTGAAG AGACGTTTGG CTGA
Start codon is in green and stop codon is in red

Lambda Repressor (CI) with degredation tag (LVA)

Part BBa_C0051 on MIT's parts registry

  • same as CI repressor, but with degredation tag on the N terminus to promote degredation

Lambda AntiRepressor (Cro)

  • Binds to OR and OL operators
  • transcriptional activator, repressor
  • favors lytic pathway in lambda bacteriophage

ATGGAACAAC GCATAACCCT GAAAGATTAT GCAATGCGCT TTGGGCAAAC CAAGACAGCT
AAAGATCTCG GCGTATATCA AAGCGCGATC AACAAGGCCA TTCATGCAGG CCGAAAGATT
TTTTTAACTA TAAACGCTGA TGGAAGCGTT TATGCGGAAG AGGTAAAGCC CTTCCCGAGT
AACAAAAAAA CAACAGCATA A
Start codon is in green and stop codon is in red

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