Summary
Four recombinant expression vectors of proinsulin including PEX2-PI, PKB49, PUC8-[Thr]6-PI and PYK10-9 were constructed using different sizes of fusion peptide genes and different regulatory genes. PYK10-9, constructed by combining the lambda PR promotor and the fused proinsulin gene containing the Lac ribosome binding site with the [Thr]6 gene, was the optimum recombinant expression vector and is estimated to give the highest yield of human proinsulin by intracellular expression in E. coli to date.
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References
Brousseau, R., Scarpulla, R., Sung, W., Hsiung, H.M., Harang, S.A., and Wu, R. (1982) Gene 17: 279–289.
Chance, R.Z., Kroeff, E.P., Hoffman, J.A. and Frank, B.H. (1981) Diabetes Care 4: 149–154.
Eun, H.M., Kang, Y., Kang, S.M., Bae, Y.S. and Yoon, J.W. (1989) Biotechniques 7: 506–510.
Frank, B.H., Pettee, J.M., Zimmerman, R.G. and Burck, P.J. (1981) In: Peptide (eds. Rich, P. H. and Gross, E.) Dierce Chemical Company, Rockford, IL, p. 729–738.
Goeddel, D.V., Kleid, D.G., Bolivar, F., Heyneker, H.L., Yansura, B.G., Crea, R., Hirose, T., Kroszewsk, A., Italcura, K. and Riggs, A.D. (1979). Proc. Natl. Acad. Sci. USA, 76: 106–110.
Goft, S.A., and Goldberg, A.L. (1985) Cell 41: 587–89.
Gubler, U. and Hoffman, B.J. (1983) Gene 25: 263–269.
Guo, L.H., Stepien, P.P., Tso, J.Y., Brousseau, R., Narang, S.A., Thomas, D.Y., and Wu, R. (1984) Gene 29: 251–255.
laemmli, U.K. (1970) Nature 227: 680–685.
Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, NY, New York, pp. 7.19–7.29.
Shen, S-H. (1984) Proc. Natl. Acad. Sci. USA. 81: 4627–4631.
Stanley, K.K. and Luzia, J.P. (1984) EMBO J. 3: 1429–1434.
Sung, W.L., Yao, F.L., Zahab, D.M., and Narang, S.A. (1986) Proc. Natl. Acad. Sci. U.S.A. 83: 561–565.
Thim, L., Hansen, M.T., Norris, K., Hoegh, I., Boel, E., Forston, J., Ammer, G., and Fiil, N. (1986) Proc. Natl. Acad. Sci. USA 83: 6766–6770.
Thim, L., Hansen, M.T., and Sorensen, A.R. (1987) FEBS Letter. 212: 307–312.
Vieira, J. and Messing, J. (1982) Gene 19: 259–268.
Wetzel, R., Kleid, D.G., Crea, R., Heyneker, H., Yansura, D.G., Hirose, T., Kraszewski, A., Riggs, A.D., Itakura, K., and Goeddel, D.V. (1981) Gene 16: 63–71.
Williams, D.C., Van Frank, R.M., Muth, W.L., and Burnett, J.P. (1982) Science 215: 687–689.
Yoon, J.W., Bachurski, C.J. and Rayfield, E.J. (1984) In: Methods in Diabetes S. L. Pohl and J. Larner editors, John Wiley and Sons Inc., New York, Vol. 1-B, pp. 165–172.
Yoon, J.W., Kang, Y. and Park, H.S. (1988) In: Recombinant DNA Techniques, J. W. Yoon, editor, KOSCO Inc., Seoul, pp. 93–115.
Yoon, J.W., Lesniak, M.A., Fussganger, R. and Notkins, A.L. (1976) Nature 264: 178–180.
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Kang, Y., Yoon, JW. Development of a high-expression vector (PYK10-9) of human proinsulin gene. Biotechnol Lett 13, 755–760 (1991). https://doi.org/10.1007/BF01088182
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DOI: https://doi.org/10.1007/BF01088182