Reference - Detail
| RRC ID | 57383 |
|---|---|
| Author | Kita H, Cho J, Matsuura T, Nakaishi T, Taniguchi I, Ichikawa T, Shima Y, Urabe I, Yomo T. |
| Title | Functional Qbeta replicase genetically fusing essential subunits EF-Ts and EF-Tu with beta-subunit. |
| Journal | J Biosci Bioeng |
| Abstract |
Qbeta replicase, an RNA-dependent RNA polymerase of RNA coliphage Qbeta, is a heterotetramer composed of a phage-encoded beta-subunit and three host-encoded proteins: the ribosomal protein S1 (alpha-subunit), EF-Tu, and EF-Ts. Several purification methods for Qbeta replicase were described previously. However, in our efforts to improve the production of Qbeta replicase, a substantial amount of the beta-subunit overproduced in Escherichia coli cells was found as insoluble aggregates. In this paper, we describe two kinds of method of producing Qbeta replicase. In one kind, both EF-Tu and EF-Ts subunits were expressed with the beta-subunit, and in the other kind, the beta-subunit was genetically fused with EF-Tu and EF-Ts. The fused protein, a single-chain alpha-less Qbeta 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. |
| Volume | 101(5) |
| Pages | 421-6 |
| Published | 2006-5-1 |
| DOI | 10.1263/jbb.101.421 |
| PII | S1389-1723(06)70604-1 |
| PMID | 16781472 |
| MeSH | Enzyme Activation Escherichia coli / genetics* Escherichia coli / metabolism* Genetic Enhancement Protein Engineering / methods* Protein Subunits Q beta Replicase / biosynthesis* Q beta Replicase / chemistry* Q beta Replicase / genetics Q beta Replicase / isolation & purification Recombinant Fusion Proteins / chemistry Recombinant Fusion Proteins / isolation & purification Recombinant Fusion Proteins / metabolism |
| IF | 2.032 |
| Times Cited | 19 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 5 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| DNA material | pET-TuTs (RDB17457) pBAD33betaH (RDB17458) pBAD33Ts-Tu-beta-3 (RDB17459) |