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Custom DNA/RNA Oligos> | o r g o n N o v e m b e r 2 0 , 2 0 1 7 Materials and methods Constructs and cell lines An oligonucleotide containing the signal sequence of human gp135/ podocalyxin (BC112035), the sequence for the SNAPf tag (from pSNAPf; New England Biolabs), and an HA epitope tag was synthe- sized and inserted into pUC57 vector (GenScript). | Get A Quote |
In polarized epithelial cells, newly synthesized cell surface proteins travel in carrier vesicles from the trans Golgi network to the apical or basolateral plasma membrane. Despite extensive research on polarized trafficking, the sites of protein delivery are not fully characterized. Here we use the SNAP tag system to examine the site of delivery of the apical glycoprotein gp135. We show that a cohort of gp135 is delivered to a ring surrounding the base of the primary cilium, followed by microtubule-dependent radial movement away from the cilium. Delivery to the periciliary ring was specific to newly synthesized and not recycling protein. A subset of this newly delivered protein traverses the basolateral membra... More