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 language | English |
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Pages (from-to) | 651-659 |
Number of pages | 9 |
Journal | Applied Biochemistry and Biotechnology |
Volume | 169 |
Issue number | 2 |
DOIs | |
Publication status | Published - Jan 2013 |