WormBase Tree Display for Construct: WBCnstr00010275
expand all nodes | collapse all nodes | view schema
| WBCnstr00010275 | Other_name | Expr1433_Ex | |
|---|---|---|---|
| Summary | [vab-8::gfp] | ||
| Driven_by_gene | WBGene00006874 | ||
| Gene | WBGene00006874 | ||
| Fusion_reporter | GFP | ||
| Type_of_construct | Translational_fusion | ||
| Construction_summary | [vab-8::gfp] translational fusions. A VAB-8SGFP reporter was constructed by cloning a 9.9 kb PstI vab-8 genomic fragment encompassing exons 613 and most of exon 14 (bp 1,71011,657) into the PstI site of p76-L18GFP This construct produces a tripartite fusion protein consisting of sequences from VAB-8, UNC-76, and GFP (VAB-8-UNC-76GFP). To generate the mec-3-vab-8L-gfp misexpression construct, full-length VAB-8L cDNA was digested with SphI and SalI and the intervening sequences were replaced with SphI( 22)-SalI( 1,037) genomic sequences. The mec-3 promoter was amplified from pPD57.56 (a gift from Andrew Fire) and placed into SacIINotI sites of pBluescript II SK(-), 9 bp upstream from the vab-8 initiator methionine. GFP was then cloned intothe BstEII site as above. The resulting mec-3-vab-8L-gfp construct was digested with PstI and religated, removing 2,038 bp that encoded exons 513 and fusing in frame the remaining portions of exons 4 and 14 to create mec-3-vab-8-gfp. Both mec-3-vab-8L-gfp and mec-3-vab-8-gfp were injected at 150 ng/ l along with rol-6 DNA into wild-type hermaphrodites to establish lines carrying these constructs. | ||
| Clone | pPD57.56 | ||
| Used_for | Transgene_construct | WBTransgene00027514 | |
| Reference | WBPaper00003058 |
