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Protein Expr Purif. 1993 Apr;4(2):160-3.

Overcoming inclusion body formation in a high-level expression system.

Author information

1
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany.

Abstract

Attempts at overexpressing T4-phage deoxycytidylate deaminase using the pET3c/BL21(DE3)/pLysS system resulted in this enzyme being part of an inactive inclusion-body complex. However, by employing an enriched growth medium it was found that the deaminase could be induced in a soluble active form to at least 20% of this organism's cellular protein. Insoluble inclusion bodies were obtained with less rich media. This procedure was employed successfully with other highly expressed proteins that formed inclusion bodies. The use of a rich growth medium during the course of protein induction may be a valuable adjunct to limiting inclusion body formation with this as well as other expression systems.

PMID:
8471848
DOI:
10.1006/prep.1993.1022
[Indexed for MEDLINE]

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