| Primary Identifier | Expr2852 | Reporter Gene | Two GFP::POP transgenes under control of a promoter that drives expression at the same apparent level in many cells, including Z1, Z4, and their descendants. One transgene, GFP::POP(del1-5), has GFP-coding sequences fused in frame to the sixth codon of pop-1 cDNA, while the other, GFP::POP(FL), fuses GFP to the full-length pop-1 cDNA. Both transgenes displayed similar expression levels and response to Wnt/MAPK signaling. In addition, both transgenes exhibited similar POP-1 asymmetry in the Z1/Z4 daughters. GFP::POP(del1-5) had dominant negative activity and was not viable in certain mutant backgrounds; by contrast, GFP::POP(FL) had only marginal dominant negative effects and rescued a pop-1 mutant. [jmp#1::pop-1-gfp] translational fusion constructs. Two GFP::POP-1 constructs, GFP::POP-1(del1-5) and GFP::POP-1(FL), were made with GFP fused at the N terminus of the pop-1 cDNA. These reporters were placed under control of a promoter expressed in Z1 and Z4 as well as many other tissues, called jmp#1. GFP::POP-1(del1-5) was made by first amplifying GFP with primers containing SacI sites at the 5 ends. This GFP fragment was cloned in frame into the SacI site of pJK706, which inserts GFP upstream of amino acid 6 of the POP-1 protein. GFP::POP-1(del1-5) was subcloned into pPD49.26 to add the unc-54 3'-UTR and then cloned into pDPMM0166 to add the unc-119 gene for use as a selective marker when producing transgenics. The resulting plasmid is named pJK789. The second construct, POP-1::GFP(FL), differs from the first only in that GFP is fused in frame upstream of the first methionine of the pop-1 cDNA. This construct is called pJK908. |
| Subcellular Localization | The developing hypodermis and the early gonad was observed using the GFP::POP reporters. As seen in previous studies, GFP was more abundant in anterior than in posterior daughters in hypodermal lineages. Expression in the early gonad departed from this anterior-posterior asymmetry: the nucleus of Z1.p was brighter than that of Z1.a, and the nucleus of Z4.a was brighter than that of Z4.p. Proximal daughters was noted contained nuclear GFP puncta similar to those described in the anterior daughters of asymmetric divisions in the embryo. |
| Definition | Name | Synonym | Primary Identifier |
|---|---|---|---|
| Epidermal layer. | hypodermis | epidermis | WBbt:0005733 |
| organ producing either sperm or ova. | gonad | WBbt:0005175 |
| WormBase Gene ID | Gene Name | Sequence Name | Organism |
|---|---|---|---|
| WBGene00004077 | pop-1 | W10C8.2 | Caenorhabditis elegans |
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