Facile production and rapid purification of functional recombinant Qβ replicase heterotetramer complex

Karthikeyan Gunasekaran, Peter L. Bergquist, Anwar Sunna*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

We describe an improved method for the production of recombinant Qβ replicase heterotetramer. The successful expression of the soluble Qβ RNA polymerase complex depends on the EF-Ts and EF-Tu subunits being co-expressed prior to β-subunit expression. Efficient co-expression requires two different inducible operons to co-ordinate the expression of the heterotrimer. The complete heterotetramer enzyme complex is achieved by production of the recombinant S1-subunit of Qβ replicase in a separate host. This approach represents a facile way for producing and purifying large amounts of soluble and active recombinant Qβ replicase tetramer without the necessity of a His-tag for purification.

Original languageEnglish
Pages (from-to)651-659
Number of pages9
JournalApplied Biochemistry and Biotechnology
Volume169
Issue number2
DOIs
Publication statusPublished - Jan 2013

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