WormBase Tree Display for Construct: WBCnstr00018781
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| WBCnstr00018781 | Other_name | Expr11335_Ex | |
|---|---|---|---|
| Summary | [ges-1.WGATAR::pJM77] | ||
| Sequence_feature | WBsf919554 | ||
| Gene | WBGene00001578 | ||
| Construction_summary | Six or seven copies of WGATAR sites in either orientation were inserted into the test vector pJM77. The vector pJM77 used to test the enhancer activity of candidate sequences was constructed as follows: a 446-bp Sau3A fragment from the promoter of the C. elegans heat shock gene 16-48 was isolated from plasmid pPC16.48-1 (Stringham et al., 1992) and inserted in the correct orientation into BamHI-cleaved vector pPD96.04 (kindly provided by A. Fire, Carnegie Institute of Washington, Baltimore,MD). In this construct, the heat shock elements of the 16-48 gene are intact but can be removed either by PstI digestion or by double digestion with PstI and HindIII. pJM77 contains the transcription initiation site, the 5'-UTR, the ATG codon, and the first 15 amino acids of the 16-48 heat shock protein fused to a GFP-lacZ reporter incorporating 15 synthetic introns. Sequence elements to be tested for enhancer activity are first multimerized, cloned into the EcoRV site of pBluescript, and transferred as a HindIII-PstI fragment into HindIII-PstI-cleaved pJM77, thereby removing the original heatshock elements and preserving insert orientation. | ||
| Used_for | Transgene_construct | WBTransgene00031454 | |
| Reference | WBPaper00004994 | ||
| Remark | pJM77 |
