Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Enzymatic production of a protein or polypeptide
Patent
1984-04-30
1987-06-16
Wiseman, Thomas S.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Enzymatic production of a protein or polypeptide
435 70, 4351723, 435253, 435317, 536 27, 935 42, 935 43, 935 73, C12P 2100, C12N 1500, C12N 120, C12N 100, C07H 1512
Patent
active
046736409
ABSTRACT:
In vivo regulation of protein production is achieved by rearranging DNA segments comprising a protein-producing gene (i.e., protein-encoding DNA as well as regulatory DNA to effect the expression of the protein-encoding DNA in the host), in response to a change in an environmental condition such as temperature. The rearrangement is synchronized and directional (irreversible) in members of the cell population, because it is catalyzed by a lambda phage site-specific recombination enzyme system that operates on a pair of lambda phage attachment sites to rapidly drive the rearrangement and to avoid the reverse reaction. The cells include means to produce the lambda enzyme system in response to the change in environmental condition. By engineering one of the attachment sites within the gene that produces the protein whose production is to be regulated (yielding two gene segments), the synchronized rearrangement operates to change the gene from one configuration to another. In only one of these configurations are the gene segments positioned and oriented for protein production. Specifically, the protein-producing configuration is: gene segment one-attachment site-gene segment two. The attachment site is exogeneous to the gene, i.e, it does not occur in that location naturally and is positioned there by engineering techniques. The regulated protein production is useful, e.g., in fermenting the desired product, by allowing cell growth to proceed in the absence of product formation. When the desired cell mass is achieved, product production is enabled by the rearrangement.
REFERENCES:
Murray, In Lambda II. R. Hendrix et al., eds. Cold Spring Harbor Laboratory, 1983, pp. 395-432.
Podhajska, Gene 40: 163-168 (1985).
Proceedings of the XV International Congress of Genetics (1983) New Delhi, India--Table of Contents and Author Index.
Cell, "Structural Features of Site-Specific Recombination at a Secondary att Site in galT" Bidwell et al. Bo. 16, (1979).
Cell, "A Secondary Attachment Site for Bacteriophage in trpC of E. coli" Christie et al., vol. 16, (1979).
Campbell et al., "Evolution of Lambdoid Phages" in Lambda II (Hendrix, Ed) Cold Spring Harbor 1983.
Silverman et al., (1981) Virology vol. 127:17-26.
Iino et al., "Trans-acting Genes of Bacteriophages of Pl and Mu Mediate Inversion of a Specific DNA Segment Involved in Flagellar Phase Variation of Salmonella", Cold Spring Harbor Symposium on Quantative Biology, 45:11-16, (Cold Spring Lab. N.Y., 1981).
Plasterk (1983) Virology 127:24-36.
Echols et al., "Control of Integration and Excision" Lambda II (Hendrix et al., Ed.) Cold Spring Harbor Laboratory, 1983, pp. 75-92.
BioTechnica International, Inc.
Mays Thomas D.
Wiseman Thomas S.
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