pCEHC1-5.5H/S(1)3xHA5’+Hyg3
$30.00
From Carolyn Silfow, University of Minnesota
Vector: pBluescript SK (-)
Origin: Plasmid pCEHC1-5.5H/S(1)3xHA5′ was modified to add a selectable marker gene. It was digested with ClaI and KpnI and ligated to a 1.7 kb fragment containing the aph7” gene from Streptomyces hygroscopicus (Berthold et al. 2002). The fragment was amplified from plasmid pHyg3 by using a forward primer at the 5’ end modified to include a ClaI site and a reverse primer at the 3’ end that included the KpnI site.
Insert: Wild-type copy of Cre01.g005900 (CECH1 gene), nuclear DNA, 5559bp HindIII(mutated)-SpeI fragment, 3xHA tagged at 5’ end of gene, Selectable marker gene aph7”.
Comment: This plasmid can rescue the mutant phenotype of the cehc1 mutation in strain CC-4509.
Selection in E. coli: Ampicillin
E. coli host strain: XL-1 Blue MRF’
Sun et al. Identification of a negative regulator of the transcription factor CRR1 pathway in Chlamydomonas reinhardtii The Plant Cell in press