WormBase Tree Display for Construct: WBCnstr00011082
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| WBCnstr00011082 | Other_name | Expr3027_Ex | |
|---|---|---|---|
| Summary | [OSM-1::gfp] | ||
| Gene | WBGene00003883 | ||
| Fusion_reporter | GFP | ||
| Type_of_construct | Translational_fusion | ||
| Construction_summary | A 3.1-kb osm-1 gene fragment representing the most 3 end of the osm-1 gene was PCR amplified from cosmid C15H5 using the primers 5 -ttggatcctctgcgtctcttatccc-3 and 5 -caatcccggggacaacgaaatagccaacgg-3 (SmaI site underlined; added to facilitate the in-frame ligation of osm-1 to GFP). This 3.1-kb fragment was shuttled through PCR-Blunt (Invitrogen Corp.) and ligated into pPD95.77 using BamHI and SmaI sites. Then, an SphI-BamHI 5.2-kb fragment representing the 5 end of the osm-1 gene plus 2.5 kb of upstream sequence was excised from cosmid C15H5 and added to the aforementioned pPD95.77 vector containing the 3.1-kb 3 end fragment. Finally, a BamHI-BglII 3.8-kb genomic fragment representing the middle of the osm-1 gene was added into the pPD95.77 vector containing the 5.2- and 3.1-kb fragments, fully reconstituting the entire osm-1 gene (9.7 kb) plus 2.5 kb of upstream promoter sequence. --precise ends. | ||
| Used_for | Transgene_construct | WBTransgene00011442 | |
| Reference | WBPaper00003751 | ||
| WBPaper00042072 |
