Construction of MYC-tagged Sch9p in Candida albicans by using of long primers PCR amplification
XU Qiurong
LV Quanzhen
SUI Xue
WANG Xiaojuan
YAN Lan
JIANG Yuanying
Abstract：Objective To construct the MYC?tagged Sch9p in Candida albicans. Methods Using a pair of the long primers to amplify sequences containing the MYC tag and the ARG4 selection markers from the plasmid pFA?ARG4?MYC.The amplified plasmid sequences were transformed into the C?terminus of SCH9 open reading frame (ORF) in C.albicans SN152 by homologous recombina?tion.The positive colonies were selected in the SC?Leu? selective solid culture medium.Afterwards,the positive colonies with correct integration were confirmed by PCR of genomic DNA.Finally,these positive transfectants were examined by time?growth curve testing, drug sensitivity spot assays,and mycelium inducing experiments.Results Strains with MYC?tagged Sch9p which had normal pheno?types were selected.Conclusions The MYC?tagged Sch9p of C.albicans strains can be constructed correctly by using of long primers to amplify the plasmid sequences containing the MYC tag and ARG4 selection markers which were integrated into the C?terminus of SCH9 ORF in C.albicans by homologous recombination.
Keywords：Candida albicansMYCSCH9long primer
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 129-133 )