WormBase Tree Display for Construct: WBCnstr00010504

WBCnstr00010504 Other_name: Expr1888_Ex
  Summary: [pat-4::gfp]
  Driven_by_gene: WBGene00003931
  Gene: WBGene00003931
  Fusion_reporter: GFP
  Type_of_construct: Translational_fusion
  Construction_summary: The plasmid pAT4.4 was constructed by replacing the 235-bp BstXI-EagI fragment of pAT4.3 with the 783-bp BstXI-EagI fragment of pAT4.1. A mutagenic PCR fragment, generated with primers BW190 and BW191 and corresponding to vertebrate kinase-dead (E359K) ILK, was cloned into the 6.69-kb SacI-NheI fragment of either pAT4.4 or pAT4.3 to generate pAT4.6 or pAT4.16, respectively. --precise ends.|[pat-4::gfp] translational fusion constructs. Plasmids pAT4.4, pAT4.6, pAT4.16, and pAT4.21 are pat-4::gfp minigenes in which the pat-4-coding region is fused in frame to the carboxy terminus of GFP. These minigenes either contain no pat-4 intronic sequence (pAT4.16) or contain the first intron of pat-4 (pAT4.4, pAT4.6, and pAT4.21).|Plasmid pAT4.1 was constructed by cloning a PCR fragment, generated using primers BW73 and BW59, into the 4285-bp BamHI-NcoI fragment of plasmid yk199a2 (GenBank accession number C39322). This plasmid contains 1915 bp of 5' UTR sequence and the first intron of the pat-4 gene. --precise ends.|The plasmid pAT4.3 was constructed by cloning a PCR product, generated using primers BW99 and BW100 and corresponding to full-length pat-4 cDNA, into the vector pPD118.20 digested with EcoRI. The 2.13-kb MluI-KpnI fragment of the resulting plasmid was then replaced with a PCR product generated using BW-116 and BW-118 and digested with MluI and KpnI. --precise ends.|The plasmid pAT4.21 was constructed by replacing the 368-bp SacI-NheI wild-type-coding sequence of pAT4.4 with a mutagenic PCR fragment generated with primers BW229 and BW230, corresponding to vertebrate kinase-dead (S343A) ILK. --precise ends.
  Clone: yk199a2
    pPD118.20
  Used_for: Transgene_construct: WBTransgene00027738
  Reference: WBPaper00005261