Construction of Expression Shuttle Vectors for the Haloarchaeon Natrinema sp. J7 Based on Its Chromosomal Origins of Replication

Figure 4

Determination of whether pFJ1, pFJ4, and pFJ6 could integrate into the J7 chromosome by Southern blot analysis. HindIII-digested total DNA extracted from cultures of J7-F clones carrying the pFJ1 (lanes 2–4), pFJ4 (lanes 6–8), or pFJ6 (lanes 10–12) plasmid were (a) separated on a 1% agarose gel at 20 V for >12 h and then (b) subjected to Southern blot analysis. The pFJ1, pFJ4, and pFJ6 plasmids were propagated in E. coli, digested with HindIII, and loaded on lanes 1, 5, and 9 as controls. The DIG-labeled pyrF sequence was used as a hybridization probe.

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