|Citation||Wehrli, M. (2003). UAS-fz2-eCFP and UAS-Myr-Arrintra. (Export to RIS)|
|Publication Type||Personal communication to FlyBase|
|Text of Personal Communication||
From wehrlim@XXXXXXXXXXXXXXX Thu Nov 06 20:47:27 2003
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
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
Please let me know if you need addtional information.
|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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