WormBase Tree Display for Transgene: WBTransgene00006877
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| WBTransgene00006877 | Public_name | syEx23 | |
|---|---|---|---|
| Summary | [Phsp-16.41::lin-3] | ||
| Synonym | (hsp::lin-3) | ||
| [pRH51 C14G10 pMOB] | |||
| Construction | Construct | WBCnstr00006714 | |
| Construction_summary | DNA encoding the EGF domain of LIN-3 from Val-142 to Pre-204 was directly amplified from a lin-3 cDNA by PCR using the primers V123 (5'-GG CAT ATG GTT CGG AAG GAA ATC GAG-3'), which adds an Ndel site and initiation codon to the 5' end of the product, and P185 (5'-T AGG ATC CTA TGG GGC ATA GAA CGC CTG-3'), which adds a termination codon and BamHI site to the 3' end. The PCR product was end-filled and subcloned into the EcoRV site of pBluescript SK(+) to create pLIN-3EGF. syEx21and syEx23 were obtained by microinjection of pRH51 at 50 ng/ul,C14G10 at 20 ng/ul, and pMOB at 30 ng/ul, into unc-31(e169) animals. The pRH51 vector contains a portion of the hsp16-41 heat shock promoter that is active in most tissues (Stringham et al., 1992).Translation should initiate in a synthetic sequence that encodes a signal peptide (Perryet al., 1993). pRH51 also contains the 3'-untranslated region of the unc-54 gene and a synthetic intron to ensure that the messenger RNA is stable and processed properly (Fire et al.,1990). pRH51 contains the insert of pLIN-3EGF subcloned into the 3' end of the synthetic signal peptide of pRH45 in the sense orientation. pRH51 was constructed by ligating an end-filled BamHI-Ndel insert fragment of pLIN-3EGF to an end-filled Ncol digest of pRH45. To create pLIN-3EGF, DNA encoding the EGF domain of LIN-3 from Val-142 to Pre-204 was directly amplified from a lin-3 cDNA by PCR using the primers V123 (5'-GG CAT ATG GTT CGG AAG GAA ATC GAG-3'), which adds an Ndel site and initiation codon to the 5' end of the product, and P185 (5'-T AGG ATC CTA TGG GGC ATA GAA CGC CTG-3'), which adds a termination codon and BamHI site to the 3' end. The PCR product was end-filled and subcloned into the EcoRV site of pBluescript SK(+). (Katz et al., 1995). | ||
| Genetic_information | Extrachromosomal | ||
| Reference | WBPaper00002043 | ||
| WBPaper00002106 | |||
| WBPaper00002216 | |||
| WBPaper00003135 | |||
| Species | Caenorhabditis elegans |
