High-level expression of bovine growth hormone in Escherichia coli results in the formation of distinct cytoplasmic granules that are visible with the phase-contrast microscope. These granules were examined by transmission and scanning electron microscopy. Intact granules were isolated from crude cell lysates by differential centrifugation and were further purified by a simple washing procedure that yields nearly homogeneous bovine growth hormone.
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Gribskov, M. and Burgess, R.R. 1983. Overexpression and purification of the sigma subunit of Escherichia coli RNA polymerase. Gene 26: 109–118.
Masui, Y., Mizuno, T. and Inouye, M. 1984. Novel high-level expression cloning vehicles: 104-fold amplification of Escherichia coli minor protein. Bio/Technology 2 81–85.
Ho, Y.-S., Lewis, M. and Rosenberg, M. 1982. Purification and properties of a transcriptional activator. J. Biol. Chem. 257: 9128–9134.
Simons, G., Remaut, E., Allet, B., Devos, R. and Fiers, W. 1984. High-level expression of human interferon gamma in Escherichia coli under control of the PL promoter of bacteriophage lambda. Gene 28 55–64.
Devos, R., Plaetinck, G., Cheroutre, H., Simons, G., Degrave, W., Tavernier, J., Remaut, E. and Fiers, W. 1983. Molecular cloning of human intereukin 2 cDNA and its expression in E. coli. Nucleic Acids Res. 11: 4307–4323.
Cheng, Y.-S.E., Kwoh, D.Y., Kwoh, T.J., Soltvedt, B.C. and Zipser, D. 1981. Stabilization of a degradable protein by its Overexpression in Escherichia coli. Gene 14: 121–130.
Itakura, K., Hirose, T., Crea, R., Riggs, A.D., Heyneker, H.L., Bolivar, F. and Boyer, H.W. 1977. Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin. Science 198: 1056–1063.
Kleid, D.G., Yansura, D., Small, B., Dowbenko, D., Moore, D.M., Grubman, M.J., McKercher, P.D., Morgan, D.O., Robertson, B.H. and Bachrach, H.L. 1981. Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine. Science 214: 1125–1129.
Williams, D.C., Van Frank, R.M., Muth, W.L. and Burnett, J.P. 1982. Cytoplasmic inclusion bodies in Escherichia coli producing biosynthetic human insulin proteins. Science 215: 687–689.
Paul, D.C., Van Frank, R.M., Muth, W.L., Ross, J.W. and Williams, D.C. 1983. Immunocytochemical demonstration of human proinsulin chimeric polypeptide within cytoplasmic inclusion bodies of Escherichia coli. European J. Cell Biol. 31: 171–174.
Schoner, B.E., Hsiung, H.M., Belagaje, R.M., Mayne, N.G. and Schoner, R.G. 1984. Role of mRNA translational efficiency in bovine growth hormone expression in Escherichia coli. Proc. Natl. Acad. Sci. (USA) 81: 5403–5407.
Prouty, W.F. and Goldberg, A.L. 1972. Fate of abnormal proteins in E. coli accumulation in intracellular granules before catabolism. Nature New Biol. 240: 147–150.
Prouty, W.F., Karnovsky, M.J. and Goldberg, A.L. 1975. Degradation of abnormal proteins in Escherichia coli. J. Biol. Chem. 250: 1112–1123.
Millonig, G. 1961. Advantages of a phosphate buffer for osmium tetraoxide solutions in fixation. J. Appl. Phys. 32: 1637–1639.
Spurr, A.R. 1969. A low viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastructure Research 26: 31–36.
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
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Schoner, R., Ellis, L. & Schoner, B. Isolation and Purification of Protein Granules from Escherichia coli Cells Overproducing Bovine Growth Hormone. Nat Biotechnol 3, 151–154 (1985). https://doi.org/10.1038/nbt0285-151
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