Back to results
US 4,374,927 AGranted
Extrachromosomal regulation of expression
AssigneeThe Board of Trustees of the Leland Stanford Jr. University
InventorsJohn J. Sninsky, Stanley N. Cohen
Life of this patent
Estimating the term…
Abstract
Microbiological methods, compositions and transformants are used for production of organic products for controlling cellular properties. Extrachromosomal elements are used which are subject to external modulation for production of a control element. The change in the amount of production of the control element allows for enhanced expression of a gene producing a poly(amino acid) product. The change in production of the control element allowing for enhanced gene expression of the product is accompanied by amplification of the product producing gene.
Claims32
A method for producing a poly(amino acid) product on demand, said method comprising:
- (a) growing to a high density transformant microorganism host cells having at least one extrachromosomal element capable of replication in said host cells which includes:
- (1) an activatable runaway-replication vector,
- (2) components of a first regulatory system comprising a first regulatory element subject to control by an external modulator, wherein said first regulatory element regulates a gene producing a control factor,
- (3) components of a second regulatory system comprising a second regulatory element, subject to control by said control factor, wherein said second regulatory element regulates a gene producing a poly(amino acid) product,
- (4) a gene expressing a poly(amino acid) product replicated in conjunction with said runaway-replication-vector,
- (5) a gene expressing a control factor;
- (b) subjecting said cells to an external modulator which modulates expression of said control factor producing gene in a direction enhancing expression of said product producing gene, while activating said runaway-replication vector, where enhanced expression and amplification of said product producing gene occurs with enhanced production of said poly(amino acid).
4 more elements
A method for producing a poly(amino acid) product on demand, said method comprising:
- (a) growing to a high density transformant microorganism host cells having two extrachromosomal elements capable of replication in said host cell, wherein a first extrachromosomal element includes:
- (1) components of a first regulatory system comprising a first regulatory element subject to control by an external modulator, wherein said first regulatory element regulates a gene expressing a control factor;
- and
- (2) a gene expressing a control factor;
- and a second extrachromosomal element includes:
- (1) an activatable runaway-replication vector;
- (2) components of a second regulatory system comprising a second regulatory element subject to control by said control factor, wherein said second regulatory element regulates a gene expressing said poly(amino acid) product;
- and
- (3) a gene expressing said poly(amino acid) product;
- (b) subjecting said cells to said external modulator which modulates expression of said control factor producing gene in a direction enhancing expression of said product producing gene, while activating said runaway-replication vector, where enhanced expression and amplification of said product producing gene occurs with enhanced production of said poly(amino acid).
7 more elements
A microorganism cell having at least one extrachromosomal element capable of replication in said cell which includes:
- (1) an activatable runaway-replication vector;
- (2) components of a first regulatory system comprising a first regulatory element subject to control by an external modulator, wherein said first regulatory element regulates a gene producing a control factor;
- (3) components of a second regulatory system comprising a second regulatory element subject to control by said control factor, wherein said second regulatory element regulates a gene producing a poly(amino acid) product;
- (4) a gene expressing said poly(amino acid) product replicated in conjunction with said runaway-replication vector,
- (5) a gene expressing a control factor.
2 more elements
A method for producing a poly(amino acid) product on demand, said method comprising:
- (a) growing to a high density transformant microorganism host cells having at least one extrachromosomal element capable of replication in said host cells which include:
- (1) an activatable runaway-replication vector;
- (2) components of a first regulatory system comprising a first regulatory element regulating a gene producing a control factor;
- (3) a gene expressing a control factor;
- (4) components of a second regulatory system comprising a second regulatory element, subject to control by said control factor, wherein said second regulatory element regulates a gene producing a poly(amino acid) product wherein said control factor is a component of said second regularoty system;
- (5) a gene expressing a poly(amino acid) product replicated in conjunction with said runaway-replication vector;
- wherein at least one of said first and second regulatory systems is subject to control by an external modulator;
- (b) subjecting said cells to an external modulator which modulates at least one of said first and second regulatory systems in a direction enhancing expression of said product producing gene, while activating said runaway-replication vector, wherein enhanced expression and amplification of said product producing gene occurs with enhanced production of said poly(amino acid).
5 more elements
A microorganism cell having at least one extrachromosomal element capable of replication in said cell which include:
- (1) an activatable runaway-replication vector;
- (2) components of a first regulatory system comprising a first regulatory element which regulates a gene producing a control factor;
- (3) components of a second regulatory system comprising a second regulatory element subject to control by said control factor, wherein said second regulatory element regulates a gene producing a poly(amino acid) product, and said second regulatory system includes said control factor;
- (4) a gene expressing said poly(amino acid) product;
- (5) a gene expressing said control factor;
- wherein at least one component of said first and second regulatory systems is subject to control by an external modulator.
3 more elements
Dependencies are inferred from the claim text.