Rapid and efficient purification of native histidine-tagged protein expressed by recombinant vaccinia virus - PubMed (original) (raw)
Rapid and efficient purification of native histidine-tagged protein expressed by recombinant vaccinia virus
R Janknecht et al. Proc Natl Acad Sci U S A. 1991.
Abstract
Vaccinia virus has been used as a vector to express foreign genes for the production of functional and posttranslationally modified proteins. A procedure is described here that allows the rapid native purification of vaccinia-expressed proteins fused to an amino-terminal tag of six histidines. Extracts from cells infected with recombinant vaccinia virus are loaded onto Ni2+.nitrilotriacetic acid (Ni2+.NTA)-agarose and histidine-tagged proteins are selectively eluted with imidazole-containing buffers. In the case of the human serum response factor (SRF), a transcription factor involved in the regulation of the c-fos protooncogene, the vaccinia-expressed histidine-tagged SRF (SRF-6His) could be purified solely by this step to greater than 95% purity. SRF-6His was shown to resemble authentic SRF by functional criteria: it was transported to the nucleus, bound specifically the c-fos serum response element, interacted with the p62TCF protein to form a ternary complex, and stimulated in vitro transcription from the serum response element. Thus, the combination of vaccinia virus expression and affinity purification by Ni2+.NTA chromatography promises to be useful for the production of proteins in a functional and posttranslationally modified form.
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References
- J Virol. 1977 Feb;21(2):796-801 - PubMed
- New Biol. 1990 Sep;2(9):751-8 - PubMed
- Mol Cell Biol. 1991 Mar;11(3):1270-80 - PubMed
- J Virol. 1988 Jun;62(6):1849-54 - PubMed
- Proc Natl Acad Sci U S A. 1988 Oct;85(19):7206-10 - PubMed
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