Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1988-07-18
1990-05-08
Martinell, James
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C12N 900
Patent
active
049238086
ABSTRACT:
A method is provided for identifying mutants that leads to the enhanced expression and periplasmic secretion of heterologous preproteins from host cells. Mutants in the host cell or in the gene encoding the heterologous preprotein are identified readily by transforming the host cell with a fusion which comprises DNA encoding the heterologous preprotein and a detectable marker protein. A marker protein is chosen that only becomes active in the periplasmic or extracellular environment. Thus, desired mutants are identified by enhanced marker expression and secretion. The ability of the preprotein DNA or host cell mutants to enhance expression and secretion of the heterologous protein-enzyme fusions is found to correlate with similar enhancement of the expression and secretion of the heterologous protein alone and not as a fusion. Screening assays led to the identification of a preprotein silent mutation resulting in high-yield secretion that could not be explained under current theories and which provides a new method for obtaining high-yielding host-vector systems.
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Genentech Inc.
Martinell James
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