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 journalArticle

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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