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. 2018 Jul 24;2(1):31–37.

Figure 4.

Figure 4

Chromosomal disruption of RPC10. A. Restriction map of RPC10 chromosomal locus and replacement of the Nru I-EcoN I internal fragment by HIS3 gene. Restriction sites are: H, Hind III; E, EcoR I; N, Nru I; En, EcoN I; B, BamH I. The arrows indicate the size of relevant restriction fragments. B. Southern analysis of the gene disruption. The diploid strain CMY214 (his−) was transformed with the 3.7 kb EcoR I fragment containing RPC10 disrupted by the HIS3 gene. The genomic DNA of several HIS+ transformants was isolated, restricted with EcoR I, and subjected to Southern analysis by hybridization with the 2.1 kb EcoR I fragment. Lanes 1, 2, and 3 show the Southern blot from three HIS+ transformants; wt, control hybridization with DNA from the original CMY214 strain. C. Tetrad analysis from one HIS+ disrupted diploid transformant containing one wild-type and one disrupted copy of RPC10. All dissected tetrads contained only two viable spores on YPD medium which were always his−.