WormBase Tree Display for Construct: WBCnstr00006107
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| WBCnstr00006107 | Summary | [unc-37FOS::yfp, elt-2::rfp] | |
|---|---|---|---|
| Driven_by_gene | WBGene00001250 | ||
| WBGene00006773 | |||
| Fusion_reporter | RFP | ||
| YFP | |||
| Type_of_construct | Translational_fusion | ||
| Construction_summary | The unc-37 reporter gene was created utilizing l-Red-mediated recombineering in bacteria as described (Tursun et al., 2009). Briefly, the unc-37-containing fosmid (WRM0612cG05) was electroporated into E. coli strain SW105 (Warming et al., 2005). Using FLP recombinase-removable galK-based cassettes, we inserted yfp immediately preceding the stop codon at the C-terminus of unc-37, resulting in a translational fusion. Recombineered fosmids were sequenced at their recombineered junctions and correct clones were maintained in E. coli strain EPI-300 T1R (Epicentre). Fosmids were digested with SbfI and injected at 10 ng/l, together with 2 ng/l ScaI-digested rol-6(d) (pRF4; 2 ng/l) or HindIII- digested elt-2::NLS-dsRed and PvuII-digested bacterial genomic DNA (150 ng/l) to generate a complex array. The DNA was injected into wild-type N2. The resulting array is called otEx4126 (unc-37FOS::yfp). otEx4126 subsequently spontaneously integrated to generate otIs288. | ||
| Used_for | Transgene_construct | WBTransgene00006252 | |
| Reference | WBPaper00036225 |
