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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Methods Mol Biol. 2015;1276:13–29. doi: 10.1007/978-1-4939-2392-2_2

Fig. 1. Polycistronic vectors for expression of E. coli core RNAP.

Fig. 1

Representative examples of plasmids used most often are shown, other variants may be more suitable for a particular application.

A. pIA423 contains rpoA, Band C genes. An intein-chitin binding protein tag (IMPACT-Nitride;, NEB) allows for easy, one-step purification of the core RNAP on chitin matrix.

B. pVS10 (rpoA, B, C and Z) encodes a C-terminally His-tagged β′.

C. pIA900 (rpoA, B, C and Z) encodes a C-terminally His-tagged β′ which can be removed using TEV protease.

D. pIA1000 (rpoA, B, C and Z) encodes an N-terminally His-tagged β.

E. pIA1198 (rpoA, B, C and Z) encodes a β′ with C-terminal HA and His tags.

F. pIA1202 (rpoA, B, C and Z) encodes a β′ with a C-terminal minimal biotin tag (AVI tag) followed by a His tag ; the latter can be removed using TEV protease, leaving the AVI tag intact.