
Overview of Protein Expression in E. coli
Abstract
This overview unit introduces general considerations and strategies for expressing proteinsin E. coli. E. coli has two characteristics that make it ideally suited as an expression system for many kinds of proteins: it is easy to manipulate and it grows quickly in inexpensive media. These characteristics, coupled with more than 10 years' experience with expression of foreign genes, have established E. coli as the leading host organism for most scientific applications of protein expression. Despite a growing literature describing successful protein expression from cloned genes, each new gene still presents its own unique expression problems. There is no single set of methods that can guarantee successful production of every protein in a useful form. Nevertheless, the vast body of accumulated knowledge has led to a general approach that often helps to solve specific expression problems.
Figures
Videos
Literature Cited
| Literature Cited | |
| Chang, A.C.Y. and Cohen, S.N. 1978. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J. Bacteriol. 134:1141-1156. | |
| de Boer, H.A., Comstock, L.J., and Vasser, M. 1983. The tac promoter: A functional hybrid derived from the trp and lac promoters. Proc. Nat. Acad. Sci. U.S.A. 80:21-25. | |
| DeLamarter, J.F., Mermod, J.J., Liang, C.M., Eliason, J.F., and Thatcher, D.R. 1985. Recombinant murine GM-CSF from E. coli has biological activity and is neutralized by a specific antiserum. EMBO J. 4:2575-2581. | |
| Edman, J.C., Hallewell, R.A., Valenzuela, P., Goodman, H.M., and Rutter, W.J. 1981. Synthesis of Hepatitis B surface and core antigens in E. coli. Nature 291:503-506. | |
| Hamilton, C.M., Aldea, M., Washburn, B.K., Babitzke, P., and Kushner, S.R. 1989. New methods for generating deletions and gene replacements in Escherichia coli. J. Bacteriol. 171:4617-4622. | |
| Marston, F.A.O. and Hartley, D.L. 1990. Solubilization of protein aggregates. Methods Enzymol. 182:264-276. | |
| Robinson, M., Lilley, R., Little, S., Emtage, J.S., Yarranton, G., Stephens, P., Millican, A., Eaton, M., and Humphreys, G. 1984. Codon usage can affect efficiency of translation of genes in Escherichia coli. Nucl. Acids Res. 12:6663-6671. | |
| Schein, C.H. 1989. Production of soluble recombinant proteins in bacteria. Bio/Technology 7:1141-1148. | |
| Schein, C.H. and Noteborn, M.H.M. 1988. Formation of soluble recombinant proteins in Escherichia coli. is favored by lower growth temperatures. Bio/Technology 6:291-294. | |



