pML9.7-3xHA UNI2 tagged with 3xHA
$30.00
From Carolyn Silflow, University of Minnesota, September 2007
UNI2 gene tagged with 3xHA (3 copies of the 9-amino acid influenza hemagglutinin epitope)
To create pML9.7-3xHA , the pML9.7 plasmid was first modified by removing sequences between the EcoRV site and the Xba I site in the multi-cloning site positioned near the 3’ end of the UNI2 gene. This step removed the Bam HI site from the multi-cloning site. The plasmid was then digested with Bam HI which cleaves the UNI2 gene at a position 13 codons upstream of the stop codon. The Bam HI overhang was filled in using Klenow. A blunt-ended Nru I/Sma I fragment from the p3xHA plasmid was then ligated into the blunted Bam HI site.
amp resistant
Silflow CD, LaVoie M, Tam LW, Tousey S, Sanders M, Wu W, Borodovsky M, Lefebvre PA (2001) The Vfl1 Protein in Chlamydomonas localizes in a rotationally asymmetric pattern at the distal ends of the basal bodies. J Cell Biol 153:63-74 Piasecki BP, LaVoie M, Tam LW, Lefebvre PA, Silflow CD (2008) The Uni2phosphoprotein is a cell cycle regulated component of the basal body maturation pathway in Chlamydomonas reinhardtii. Mol Biol Cell 19:262-273