WormBase Tree Display for Construct: WBCnstr00006105
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| WBCnstr00006105 | Summary | [lsy-22FOS::2XFLAG::venus, elt-2::rfp] | |
|---|---|---|---|
| Driven_by_gene | WBGene00009188 | ||
| WBGene00001250 | |||
| Gene | WBGene00000584 | ||
| Fusion_reporter | Venus | ||
| Type_of_construct | Translational_fusion | ||
| Construction_summary | The lsy-22 reporter gene was created utilizing l-Red-mediated recombineering in bacteria as described (Tursun et al., 2009). Briefly, the lsy-22-containing fosmid (WRM0628cC04 )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 lsy-22, 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 otEx4125 (lsy-22FOS::2XFLAG::venus). | ||
| Used_for | Transgene_construct | WBTransgene00006250 | |
| Reference | WBPaper00036225 |
