WormBase Tree Display for Construct: WBCnstr00011571
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| WBCnstr00011571 | Other_name | Expr3648_Ex | |
|---|---|---|---|
| Summary | [ain-1::ain-1-gfp] | ||
| Driven_by_gene | WBGene00015547 | ||
| Gene | WBGene00015547 | ||
| Fusion_reporter | GFP | ||
| Type_of_construct | Translational_fusion | ||
| Construction_summary | Site-directed mutagenesis was performed on the XhoI-HpaI DNA fragment that rescued the ain-1 mutant phenotype to generate a SalI site at the stop condon. The GFP coding region from pPD103.87 was cloned into this SalI site. For the CoIP experiment, this plasmid was injected at 1 ng/ul into lin-31(n1053);ain-1(ku322) animals with the marker DNA pRF4 (100 ng/ul) that contains a rol-6 dominant mutant gene. After determining that this ain-1::gfp construct rescued the mutant phenotype, the transgene was integrated into the genome by using a standard protocol. This integrated transgene, expected to have a low copy number, was expressed at a low level. The GFP signals in the animals were barely detectable, and the protein was not detected in total protein lysate of the animals in a Western blot with an anti-GFP antibody. For expression analysis, the same ain-1::gfp constructed was injected at 10 ng/ul so that the GFP signal could be better visualized at the larval stages. | ||
| Used_for | Transgene_construct | WBTransgene00028788 | |
| Reference | WBPaper00026707 |
