ISSN:
1476-5535
Keywords:
Host strain
;
Protein stability
;
Western analysis
;
Thermoinducible high copy number replicon
;
Replicon
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Process Engineering, Biotechnology, Nutrition Technology
Notes:
Summary Murine interleukin-3 (Mu IL-3) cDNA was previously expressed inEscherichia coli using atac promoter and a constitutive high copy number plasmid vector. We found that significant increases in expression levels could be realized by using thetac promoter for the expression of Mu IL-3 in a plasmid vector possessing a temperature-inducible runaway-replicon. In contrast, use of anlpp promoter under similar conditions did not result in an increase in the Mu IL-3 expression level. Significant differences were observed when the expression levels of IL-3 were monitored in variousE. coli hosts having different genetic backgrounds. A mutant ofE. coli which lacks the protease La was found to increase the level of IL-3 produced. This report describes the effect of a specific protease-deficientE. coli host strain, as well as the effect of different promoters and plasmid replicons on the expression levels and stability of a heterologous gene product.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01569554
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