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. 2007 Jun 18;35(12):e88. doi: 10.1093/nar/gkm449

Figure 2.

Figure 2.

Analysis of S. acidocaldarius transformants. (A) Southern blot of HindIII digested genomic DNA preparations; the probes complementary to pyrE and a pRN1-fragment (see the Materials and Methods section) were used concurrently. −c: untransformed MR31, A+c: positive control: plasmid pA from E. coli, G+c: separate positive control (plasmid pG from E. coli) because of the additional HindIII restriction site present in pG. (B) Southern blot like in A for controls and cultures transformed with pC and pE after 200 generations of consecutive cultivation. (C) Colony hybridizations for S. acidocaldarius cultures transformed with pC and pE with pRN1 specific probes. (D) Restriction analysis (SacI) of retransformation experiments for all shuttle constructs, o: original plasmid prepared from E. coli, r: retransformed plasmid. (E) Restriction analysis (SacI) of shuttle vectors isolated directly from S. acidocaldarius: o: Plasmid from E. coli, S: plasmid from Sulfolobus (∼40 times more concentrated than from E. coli), r: retransformed plasmid.