WormBase Tree Display for Construct: WBCnstr00019804
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WBCnstr00019804 | Summary | [Pmyo-2::lin-18ICD::GFP] |
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Driven_by_gene | WBGene00003514 | |
Gene | WBGene00003007 | |
Fusion_reporter | GFP | |
Construction_summary | The most proximal section of the lin-18 promoter was PCR amplified using a primer containing restriction sites to be inserted (TI0013), and using SalI and BamHI restriction sites, the fragment was re-inserted into a previously generated promoter::gfp plasmid pTI02.1 (Inoue et al.,2004). This places unique NheI, MluI and BamHI sites between the 5kb promoter and the gfp coding region. PCR fragments corresponding to various sections of LIN-18 cDNA were generated by RT-PCR and inserted into pSYQ10.1. The cDNA fragment coding for the Ryk intracellular domain (ICD) was amplified using primers TI0197 and TI0018. To express LIN-18 fragments under the control of Pmyo-2, a strong pharyngeal promoter, we substituted the lin-18 promoter with the myo-2 promoter from the L4640 (myo-2::cfp)plasmid (Addgene plasmid 1662; A. Fire et al. pers. comm.). | |
Reference | WBPaper00045442 |