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Reference Report
| Reference | |||
|---|---|---|---|
| Citation | Wehrli, M. (2003). UAS-fz2-eCFP and UAS-Myr-Arrintra. (Export to RIS) | ||
| FlyBase ID | FBrf0173106 | ||
| Publication Type | Personal communication to FlyBase | ||
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| Text of Personal Communication |
From wehrlim@XXXXXXXXXXXXXXX Thu Nov 06 20:47:27 2003
Envelope-to: djs93@XXXXXXXXXXXXXXX Delivery-date: Thu, 6 Nov 2003 20:47:27 \+0000 > 1. I have been unable to trace the construct 'UAS-fz2-eCFP'. Could you> send details of its construction or point me towards a reference where> I could get these details? We made the construct by fusing the eCFP (Clontech) with its start methionine to the last amino acid of fz2 (a gift of Jeremy Nathans) while inserting an aspartate inbetween (the rationale was to provide with the aspartate a carboxy group to replace the now blocked C-terminal carboxy group). Therefore the protein sequence at the junction is (fz2)AASHV-D-MVSKGE(eCFP). The construct was assembled from restriction fragments and PCR fragments by transforming the linear fragments into yeast. The fragments were designed to have at least 20bp overlap in sequence and yeast efficiently uses these overlaps to recombine the fragments and assemble the construct. Erdeniz N, Mortensen UH, Rothstein R. Cloning-free PCR-based allele replacement methods. Genome Res. 1997 Dec;7(12):1174-83. The resulting fz2-eCFP construct was cloned into the UAS vector we previously used followed by the insertion of a white-flip-out cassette (Wehrli and Tomlinson, 1998). When the flip-out cassette is removed the UAS promoter can drive expression of the cDNA. This construct is based on those generated by Gary Struhl. The resulting construct would be UAS> GMR-white flipout cass.> fz2-eCFP \- tub 3'UTR \- Carnegie2NX > 2. What is the origin/nature of the myristoylatation tag used in the > construct 'UAS-Myr-Arrintra'? MGNKCCSKRQ (as in Struhl and Adachi, Cell, 93, 649-660) The complete protein sequence (including the 6xFLAG tags) is MGNKCCSKRQFMKSAEDYKDDDDKDYKDDDDKDYKDDDDKGSDYKDDDDKDYKDDDDKDYKDDDDKLQFCRTRIGKSRT EPKDDQATDPLSPSTLSKSQRVSKIASVADAVRMSTLNSRNSMNSYDRNHITGASSSTTNGSSMVAYPINPPPSPATRS RRPYRHYKIINQPPPPTPCSTDICDESDSNYTSKSNSNNSNGGATKHSSSSAAACLQYGYDSEPYPPPPTPRSHYHSDV RIVPESSCPPSPSSRSSTYFSPLPPPPSPVQSPSRGFT Please let me know if you need addtional information. Best wishes, Marcel |
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| Related Publication(s) | |||
| Research paper | Wg/Wnt signal can be transmitted through arrow/LRP5,6 and Axin independently of Zw3/Gsk3 activity. Tolwinski et al., 2003, Dev. Cell 4(3): 407--418 [FBrf0158859] |
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| Language of Publication | English | ||
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Data from Reference
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| Alleles (2)
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| Constructs (1)
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| Genes (2)
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