Skip to main content
. 2007 Feb 22;26(5):1279–1291. doi: 10.1038/sj.emboj.7601584

Figure 1.

Figure 1

Construction of YACs and BACs with shorter or longer alphoid DNA inserts. (A) The method for construction of deleted YACs using homologous recombination in yeast. B indicates the BamHI site. (B) Constructs were analyzed by PFGE, followed by ethidium bromide staining. Gel-purified YAC DNA was analyzed in lanes 2, 4, 6 and 8. Lanes 1 and 2, α7C5htel; lanes 3 and 4, del.24; lanes 5 and 6, del.20; lanes 7 and 8, del.22; M indicates DNA size marker. (C) HT1080 genomic DNA transformed with the YAC constructs were digested with BamHI and analyzed by PFGE, followed by Southern blot using a 32P-labeled 11-mer probe. The arrowheads indicate input alphoid DNAs. Lane 1, HT1080; lane 2, del.20HT1–5; lane 3, del.22HT1–3. (D) BACs with alphoid DNA with wild-type or mutant CENP-B boxes were constructed as shown. H, N and S indicate HindIII, NheI and SpeI sites, respectively. (E) BACs were digested with NheI and SpeI and analyzed by PFGE, followed by ethidium bromide staining. Lane 1, pWTR11.32; lane 2, pWTR11.64; lane 3, pWTR11.128; lane 4, pCBB11.1.32. M indicates DNA size marker.