WormBase Tree Display for Construct: WBCnstr00020998
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| WBCnstr00020998 | Public_name | SC-84 | |
|---|---|---|---|
| Summary | [Pgrp-1::gfp::grp-1] | ||
| Driven_by_gene | WBGene00001743 | ||
| Gene | WBGene00001743 | ||
| Fusion_reporter | GFP | ||
| Construction_summary | To construct Pgrp-1::gfp::grp-1 (SC-84), we first amplified from bGG836 a 770 bp DNA fragment containing the grp-1 promoter (the intergenic region between grp-1 and the gene immediately upstream) using primers grp1-3 [aaCGGCCGcatcggttctggaaagcttatt] and grp1-4[aaGGTACCatttatcccttgtttttccgc]. The 2.3 kb grp-1 genomic region was amplified using primers grp1-1 [aaGCTAGCatgtcatcgcggtattcagg] and grp1-2 [aaTCCGGAtcagtgggtactttttggtc], which contain NheI and BspEI sites, respectively. Both PCR products were TOPO cloned to generate plasmids SC-65(grp-1 promoter) and SC-69 (grp-1 ORF). The grp-1 promoter was then subcloned into the GFP vector pPD118.15 (Addgene, Cambridge, MA)upstream of the sequences encoding GFP using EagI and Acc65I. The grp-1 genomic region was subcloned into the resulting plasmid downstream and in frame with GFP using NheI and BspEI to generate plasmid SC-84. | ||
| Used_for | Transgene_construct | WBTransgene00021189 | |
| Reference | WBPaper00045516 |
