WormBase Tree Display for Construct: WBCnstr00014992
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| WBCnstr00014992 | Other_name | Expr9987_Ex | |
|---|---|---|---|
| Summary | [Pddo-2::GFP] | ||
| Driven_by_gene | WBGene00017565 | ||
| Fusion_reporter | GFP | ||
| Construction_summary | To generate the ddo-2 promoter-GFP fusion construct, approximately 2.6 kb genomic regions upstream of the ddo-2 initiation codon and the adjacent first exons were amplified by PCR. The template used was genomic DNA for Pddo-2::GFP. The primers used were 5'-CTG CAG CAA GAA GAA GAA GAA CGG AG-3' (forward primer) and 5'-GAG CTC GCC ATT TTA TTC TG-3' (reverse primer). The nested PCR method was employed using the following first pair of primers: 5'-ATT GAG GAG TGC ATG TGT GCT CCA C-3' (forward primer) and 5'-AAT CCT GCT GGT CCT GCA CTG CAC G-3' (reverse primer). The PCR products were cloned into a pT7Blue vector (Novagen, Madison, WI, USA), which generated pT7-Pddo-2 and the sequences were confirmed. Subsequently, the PstI-SacI fragments containing the ddo-2 promoter regions of pT7-Pddo-2 were subcloned into pPD95.67, resulting in the Pddo-2::GFP reporter plasmid. | ||
| Used_for | Transgene_construct | WBTransgene00015401 | |
| Reference | WBPaper00040844 |
