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

Figure 5

Relative copy numbers of the pFJ1, pFJ4, and pFJ6 plasmids in J7-F cells. (a) Schematic diagrams of the J7 chromosome and indicated vectors. The specific locations of Sau3AI sites in the J7 chromosome and plasmids are shown. The 200 bp 3916 pro segment, which was used as a DIG-labeled probe, is indicated with arrows and asterisks. (b) The copy numbers of pFJ1-pro3916, pFJ4-pro3916, and pFJ6-pro3916 were determined by Southern blot analysis. Sau3AI-digested total DNA from J7-F cells transformed with pFJ1 (lanes 2–4), pFJ4 (lanes 6–8), or pFJ6 (lanes 10–12) was hybridized with the DIG-labeled 3916 pro sequence. Total DNA samples from J7-F cells (lane 0), pFJ1-pro3916 (lane 1), pFJ4-pro3916 (lane 5), and pFJ6-pro3916 (lane 9) plasmids propagated in E. coli were used as controls.

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