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Attempts to engineer an Escherichia coli DNA-binding protein as a tool for affinity purification of heterologous proteins

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Biotechnology Techniques

Summary

Restriction sites were introduced into the Escherichia coli melR gene that facilitated the fusion of other proteins to MelR. Both β-galactosidase and the constant domain of the human IgG kappa light chain were fused to MelR. However, whilst unmodified MelR could be over-expressed, neither MelR fusion protein was over-produced. Addition of an extra domain to MelR leads to reduced expression in a number of genetic backgrounds.

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References

  • Bingham, A. & Busby, S. (1987) Mol. Microbiol. 1, 117–124.

    Google Scholar 

  • Casadaban, M.J. & Cohen, S.N. (1980) J. Mol. Biol. 138, 179–207.

    Google Scholar 

  • Caswell, R., Williams, J., Lyddiatt, A. & Busby, S. (1992a) Biochem. J. 287, 493–499.

    Google Scholar 

  • Caswell, R., Webster, C. & Busby, S. (1992b) Biochem. J. 287, 501–508.

    Google Scholar 

  • Caulcott, C.A. & Rhodes, M. (1986) TIBTECH 4, 142–146.

    Google Scholar 

  • Downham, M., Busby, S., Jefferis, R. & Lyddiatt, A. (1992) J. Chromatography (Biomed. App.) 584, 59–68.

    Google Scholar 

  • Gallegos, M-T., Michan, C. & Ramos, J-L. (1993) Nucl Acids Res., in press.

  • Harrison, S.C. (1991) Nature 353 715–719.

    Google Scholar 

  • Hammond, P.M, Atkinson, T., Sherwood, R.F. & Scawen, M.D. (1991) Pharm. Tech. Int. 4, 24–32.

    Google Scholar 

  • Ramos, J., Rojo, F., Zhou, L. & Timmis, K. (1990) Nucl. Acids Res. 18, 2149–2152.

    Google Scholar 

  • Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular Cloning. A Laboratory Manual; 2nd Edition. Cold Spring Harbor, NY, USA.

  • Sassenfeld, H.M. (1990) TIBTECH 8, 88–93.

    Google Scholar 

  • Schauder, B., Blocker, H., Frank, R. & McCarthy, J.E.G. (1987) Gene 52, 279–283.

    Google Scholar 

  • Schleif, R. (1988) Science 241, 1182–1187.

    Google Scholar 

  • Tobin, J. & Schleif, R. (1990) J. Mol. Biol. 211, 75–89.

    Google Scholar 

  • Uhlen, M. & Moks, T. (1990) Methods in Enzymology 185, 129–143.

    Google Scholar 

  • Ullmann, A. (1984) Gene 29, 27–31.

    Google Scholar 

  • Whittle, N., Adair, J., Lloyd, C., Jenkins, L., Devine, J., Schlom, J., Raubitschek, A., Colcher, D. & Bodmer, M. (1987) Prot. Eng. 1, 499–505.

    Google Scholar 

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Authors and Affiliations

  1. School of Biochemistry, University of Birmingham, P.O. Box 363, B15 2TT, Edgbaston, Birmingham, UK

    R. Caswell & S. Busby

  2. School of Chemical Engineering, University of Birmingham, P.O. Box 363, B15 2TT, Edgbaston, Birmingham, UK

    A. Lyddiatt & M. Downham

Authors
  1. R. Caswell
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  2. A. Lyddiatt
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  3. S. Busby
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  4. M. Downham
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Caswell, R., Lyddiatt, A., Busby, S. et al. Attempts to engineer an Escherichia coli DNA-binding protein as a tool for affinity purification of heterologous proteins. Biotechnol Tech 7, 307–312 (1993). https://doi.org/10.1007/BF00150904

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