אסיף מאגר המחקר החקלאי

Screening for highly active plasmid promoters via fusion to β-galactosidase gene

Year:

1982

Source of publication :

Authors :

Volume :

37

Co-Authors:

Rosner, A., Departments of Virology, Weizmann Institute of Science, 76100, Rehovot, Israel
Aviv, H., Departments of Virology, Weizmann Institute of Science, 76100, Rehovot, Israel
Gorecki, M., Organic Chemistry, Weizmann Institute of Science, 76100, Rehovot, Israel

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Abstract:

A plasmid containing promoter-deleted inactive β-galactosidase gene [1] was used to select promoters of the pEP121 plasmid [2]. Colonies of cells harboring reactivated ß-galactosidase gene were identified by their redcolor on McConkey plates. The quantitative amounts of β-galactosidase produced in each clone were estimated by assaying enzyme activity and by measuring the specific β-galactosidase protein following fractionation of total cells’ proteins on polyacrylamide gel. A wide range of enzyme activities was observed. The most active promoter isolated was shown to promote β-galactosidase production more efficiently, compared with the original β-galactosidasepromoter, amounting to 20% of all cell proteins. Such highly active promoters may be utilized in the future, to promote expression of cloned genes in bacteria. © 1982, Walter de Gruyter. All rights reserved.

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