Neocentromeres Form Efficiently at Multiple Possible Loci in Candida albicans
Figure 6
Chromatin immunoprecipitation (ChIP) of CENP-ACse4p detects centromere DNA.
A. CENP-ACse4p antibody specifically recognizes C. albicans centromeric DNA. Chromatin immunoprecipitation (ChIP) of wild-type strain SC5314. PCR amplification of CEN4, CEN5, CEN6 or non-centromeric TAC1 DNA from wild-type strain SC5314. Lysates (lys) were precipitated with (+) or without (−) CENP-ACse4p antibody. B. CENP-ACse4p antibody precipitates more URA3 in FOAR cells than in Ura+ cells. PCR amplification after chromatin immunoprecipitation (ChIP) was performed for Class A (YJB9907) and Class B (YJB9929) cen5Δ::URA3 strains using primers that amplify the URA3 gene that replaced the CEN5 DNA sequences. In Class B strains, URA3 did not amplify from the FOAR lysate because the cen5Δ::URA3 Chr5 homolog had been lost (Figure 4B). Numbers indicate the intensity of the +antibody (+) band relative to the lysate band for each sample.