CC-5502 d1blic::D1bLIC-crCherry KAP-GFP mt- [C5]
$30.00
From Mary Porter, University of Minnesota, March 2019
The original T8D9 background strain contains an insertion in the D1bLIC gene on chromosome 9 that results in defective retrograde IFT and short flagella. This strain contains two transgenes, D1bLIC-crCherry and KAP-GFP.
Background: CC-4487 d1blic [T8D9]
Origin: The D1bLIC-crCherry strain was co-transformed with pHyg3 and KAP-GFP, selected on hygromycin, and screened for the presence of KAP-GFP by fluorescence microscopy
Culture maintenance: Well-flagellated cells require culture in dilute media with rocking or aeration for either 2-4 hours in M-N/5 or overnight in TAP
Comment: The host D1bLIC rescued cells are paralyzed with some wiggling due to the second, uncharacterized mutation in the original T8D9 strain. The KAP-GFP transgene is expressed at the same level as the endogenous wild-type KAP protein.
Mueller J, Perrone CA, Bower R, Cole DG, Porter ME (2005) The FLA3 KAP subunit is required for localization of kinesin-2 to the site of flagellar assembly and processive anterograde intraflagellar transport. Mol Biol Cell. 16:1341-54
Chien A, Shih SM, Bower R, Tritschler D, Porter ME, Yildiz A (2017) Dynamics of the IFT machinery at the ciliary tip. Elife. Sep 20;6