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US 2003/0157661 A1Published application
De novo synthesized plasmid, methods of making and use thereof
AssigneeExpression Technologies Inc.
InventorChuan Li
Life of this patent
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Abstract
The invention relates to a de novo synthesized plasmid. The plasmid comprises relevant sequences for plasmid replication and plasmid selection. The methods of making and use of the plasmid are disclosed. The plasmid can be used to make other plasmids. These plasmids and their host cells can be used for biomedical applications.
Claims15
A de novo synthesized plasmid comprising at least a replication origin and a selection marker gene wherein;
- (a) the replication origin contains sequences relevant to autonomous plasmid replication in a host cell;
- and
- (b) the selection marker gene contains sequences relevant to the selection of a plasmid in a host cell.
A method of preparing a de novo synthesized plasmid combined from at least two DNA fragments comprising:
- (a) preparing a linear replication origin DNA fragment;
- (b) preparing a linear selection marker gene DNA fragment;
- (c) combining the DNA fragments prepared from steps (a) and (b) to form a circular de novo synthesized plasmid;
- (d) introducing the plasmid made from step (c) into a host cell;
- and
- (e) selecting the plasmid with appropriate replication origin and selection marker from transformed host cells.
3 more elements
A method of using a de novo synthesized plasmid comprising:
- (a) Linearizing the de novo synthesized plasmid;
- (b) inserting one or more functional DNA fragments to the linearized plasmid to make other plasmids;
- (c) introducing the plasmids made from step (b) into host cells;
- (d) selecting the plasmids and host cells with desired properties;
- and
- (e) using the plasmids and host cells for biomedical applications.
3 more elements
Dependencies are inferred from the claim text.