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Functional Qβ replicase genetically fusing essential subunits EF-Ts and EF-Tu with β-subunit

Paper ID Volume ID Publish Year Pages File Format Full-Text
22436 43278 2006 6 PDF Available
Title
Functional Qβ replicase genetically fusing essential subunits EF-Ts and EF-Tu with β-subunit
Abstract

Qβ replicase, an RNA-dependent RNA polymerase of RNA coliphage Qβ, is a heterotetramer composed of a phage-encoded β-subunit and three host-encoded proteins: the ribosomal protein S1 (α-subunit), EF-Tu, and EF-Ts. Several purification methods for Qβ replicase were described previously. However, in our efforts to improve the production of Qβ replicase, a substantial amount of the β-subunit overproduced in Escherichia coli cells was found as insoluble aggregates. In this paper, we describe two kinds of method of producing Qβ replicase. In one kind, both EF-Tu and EF-Ts subunits were expressed with the β-subunit, and in the other kind, the β-subunit was genetically fused with EF-Tu and EF-Ts. The fused protein, a single-chain α-less Qβ replicase, was mostly found in the soluble fraction and could be readily purified. These results pave the way for the large-scale production of the highly purified form of this enzyme.

Keywords
Qβ replicase; fusion protein; RNA amplification; RNA replication
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 101, Issue 5, May 2006, Pages 421–426
Authors
, , , , , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us