WormBase Tree Display for Construct: WBCnstr00021037
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| WBCnstr00021037 | Public_name | pPC113 | |
|---|---|---|---|
| Summary | [Peat-4::unc-7(A59T)::YFP] | ||
| Driven_by_gene | WBGene00001135 | ||
| Gene | WBGene00006747 | ||
| Fusion_reporter | YFP | ||
| Construction_summary | To drive unc-7::YFP expression from different promoters, pPC87 was cut with AfeIII and ligated with the Gateway Vector Conversion Reading frame Cassette B (Invitrogen) to generate the destination vector pPC89. The LR sites flanking unc-7 were removed using single site mutagenesis to obtain the pPC89R plasmid. The A59T mutation in the first unc-7 exon was introduced into pPC89R via single site mutagenesis to generate pPC99. To generate the minigene unc-7(A59T) version, changes were made using single site mutagenesis: two large intronic regions (intron 2 and 5) were removed from the pPC89R plasmid to generate pPC94. Subsequently, theA59T mutation was introduced into pPC94 to generate pPC104. To drive unc-7(A59Tminigene)::YFP expression from the glutamatergic neuronal promoter (Peat-4 ) the pPC52, was recombined into pPC104 using LR clonase to make pPC113 [Peat-4: unc-7(A59T)::YFP], | ||
| Used_for | Transgene_construct | WBTransgene00021232 |
