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WormBase Tree Display for Expr_pattern: Expr2646

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Name Class

Expr2646Expression_ofGeneWBGene00004804
Reflects_endogenous_expression_ofWBGene00004804
Expression_dataLife_stageWBls:0000024
WBls:0000038
WBls:0000027
WBls:0000035
WBls:0000041
WBls:0000015
WBls:0000021
WBls:0000010
WBls:0000013
Anatomy_termWBbt:0003887Certain
WBbt:0003888Certain
WBbt:0005733Certain
WBbt:0005772Certain
GO_termGO:0005634
Subcellular_localizationnuclei
TypeReporter_gene[skn-1::gfp] translational fusion. skn-1 gfp promoter fusion construct (SknPro GFP) was created by ligating GFP vector pPD95.67 and a PCR-amplified 2.1-kb clone containing the promoter region and 38 amino acids from the first ATG codon of the skn-1 gene from cosmid T19E7. To generate the SKN-1 GFP translational fusion construct, the 5.7-kb EcoRI DNA fragment that rescues the maternal skn-1 phenotype and encodes the 533-amino-acid SKN-1 protein was amplified from cosmid B0547. A ClaI site was created immediately 3' to the SKN-1 C terminus by the QuikChange method (Stratagene), which was used for all site-directed mutagenesis. This EcoRI fragment was subcloned into pUC18, which contained the upstream 1.3-kb SphIEcoRI fragment from SknPro GFP. A 0.8-kb ClaI fragment that contained the GFP open reading frame (amplified from plasmid pPD114.35) was then cloned into the ClaI site to generate an in-frame exon fusion of GFP to the SKN-1 C terminus.
PatternIn larvae and young adults, SKN-1 GFP was usually present at very low levels in intestinal nuclei. SKN-1 GFP was readily detectable in the ASI neurons, but not in other cells in the head.
In late-stage embryos, SKN-1 GFP was also present in intestinal nuclei but not in the hypodermis.
Nuclear SKN-1 GFP was uniformly detected in intestinal precursors beginning at the 50100-cell stage, then in both the intestine and hypodermis.
PictureWBPicture0000008452
RemarkGene_regulation: A promoter fusion transgene in which only the SKN-1 N terminus was linked to GFP (SknPro GFP) was constitutively expressed at high levels in all intestinal cells, suggesting that SKN-1 expression or localization might also be regulated posttranscriptionally by oxidative stress.
Gene_regulation: After exposure to either paraquat or heat, neither the location nor intensity of SKN-1 GFP was detectably altered in the ASI neurons, but in a high percentage of animals, elevated levels of SKN-1 GFP appeared in intestinal cell nuclei, particularly anteriorly and posteriorly, where GCS-1 GFP is most robustly expressed. SKN-1 GFP accumulated in intestinal nuclei within 5 min after treatment with 50 mM sodium azide, which induces oxidative stress by blocking mitochondrial electron transport.
ReferenceWBPaper00006024
TransgeneWBTransgene00000658
WBTransgene00000657