WormBase Tree Display for Transgene: WBTransgene00006675

WBTransgene00006675 Public_name: muEx108
  Summary: [pKL99-2(daf-16::GFP/daf16bKO) + pRF4(rol-6)]
  Synonym: [DAF-16::GFP]
    [Pdaf-16::DAF-16::GFP]
    [daf-16a::GFP/daf-16bKO, pRF4(rol-6(su1006))]
    [pKL99-2(DAF-6::GFP/daf16bKO)+pRF4(rol-6(su1006)]
    [pKL99-2(daf-16::GFP;rol-6(d))]
  Construction: Construct: WBCnstr00006519
    Coinjection: WBCnstr00004720
  Construction_summary: To generate this daf-16 tissue-specific promoter fusion, HindIII and SnaB1 sites were introduced immediately 5' to the first ATG by PCR amplification of the daf-16 cDNA construct (yk13f11, identified by Y. Kohara) using an inverse PCR strategy (HindIII is more proximal to the ATG). A HindIII GFP fragment from the L2911 plasmid (gift from A. Fire) was inserted into the HindIII site to generate an in-frame fusion of GFP immediately upstream of the N terminus of DAF-16a (pNL205). The SnaB1 site was used to insert the Pdaf-16, which contains 6 kb of the daf-16 5' regulatory sequence amplified by PCR from cosmid R13H8 (pNL209) (Libina et al. 2003).
    A frameshift mutation was introduced into the DAF-16b-specific exon upstream of the winged-helix domain by digestion with NgoMIV followed by 'fill-in' and religation of DAF-16a::GFP to generate DAF-16a::GFP/bKO (Lin et al. 2001). Clone = pKL99-2.
  Genetic_information: Extrachromosomal
  Associated_with: Strain: WBStrain00004850
  Reference: WBPaper00004702
    WBPaper00028865
    WBPaper00035871
  Species: Caenorhabditis elegans