Linear and circular expression elements
AssigneeBoard of Regents, The University of Texas System
InventorsKathryn F. Sykes, Stephen Albert Johnston
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Abstract
The present invention relates to linear expression elements (LEEs) and circular expression elements (CEEs), which are useful in a variety of molecular biology protocols. Specifically, the invention relates to the use of LEEs and CEEs to screen for gene function, biological effects of gene function, antigens, and promoter function. The invention also provides methods of producing proteins, antibodies, antigens, and vaccines. Also, the invention relates to methods of making LEEs and CEEs, and LEEs and CEEs produced by such methods.
Claims96
A method of assaying for the production or regulation of expression of at least one polypeptide from a linear or circular nucleic acid segment comprising a promoter or putative promoter and an open reading frame or putative open reading frame, the method comprising:
- a) obtaining at least one linear or circular nucleic acid segment comprising a promoter or putative promoter and an open reading frame encoding a peptide or putative open reading frame;
- b) placing the nucleic acid segment under conditions conducive to expression of the polypeptide from the open reading frame;
- and
- c) assaying for the production or regulation of expression of a polypeptide from the open reading frame or putative open reading frame.
A method of analyzing a nucleic acid sequence comprising:
- a) obtaining a nucleic acid segment;
- b) linking the nucleic acid segment to a promoter and a terminator to create a linear or circular expression element;
- c) providing the linear or circular expression element to a cell-free expression system or to a cell under conditions conducive to expression of any product encoded for by the nucleic acid segment;
- and
- d) analyzing any expression of any product encoded by the nucleic acid sequence.
A method of analyzing a nucleic acid segment for activity as a promoter comprising:
- a) obtaining a nucleic acid segment encoding a putative promoter;
- b) linking the nucleic acid segment encoding the putative promoter to a nucleic acid encoding a polypeptide to create a linear or circular expression element;
- c) providing the linear or circular expression element to a cell-free expression system or to a cell under conditions conducive to expression of the polypeptide;
- and
- d) analyzing any expression of the polypeptide.
A method of screening for a biological response comprising:
- a) obtaining a linear or circular expression element by a process comprising:
- obtaining a DNA segment comprising an open reading frame;
- linking the open reading frame to a promoter and a terminator to create a linear or circular expression element;
- and
- b) providing the expression element to an organism under conditions conducive to expression of any product encoded for by the open reading frame.
A method of vaccinating an organism comprising:
- a) obtaining a linear or circular expression element by a process comprising obtaining a DNA segment comprising an open reading frame and linking the open reading frame to a promoter and a terminator to create a linear expression element;
- and
- b) providing the linear expression element to an organism under conditions conducive to expression of any product encoded for by the open reading frame, such that immune response is produced in the animal.
A method of selecting open reading frames effective for generating an immune response specific to a pathogen in an organism, comprising:
- a) preparing a plurality of linear or circular expression elements produced by a method comprising:
- obtaining plurality of DNA segments comprising open reading frames from a pathogen;
- and
- linking open reading frames to promoters and terminators to create a plurality of linear or circular expression elements;
- b) introducing the plurality of linear or circular expression elements into an organism;
- and
- c) selecting from the plurality of linear or circular expression elements open reading frames that are effective to generate said immune response.
A method of producing a linear or circular expression element comprising:
- a) obtaining a DNA segment comprising an open reading frame;
- and
- b) linking the open reading frame to a promoter and a terminator to create a linear or circular expression element.
A linear or circular expression element produced by a method comprising:
- a) obtaining a DNA segment comprising an open reading frame;
- and
- b) linking the open reading frame to a promoter and a terminator to create a linear or circular expression element.
A linear or circular expression element comprising a DNA segment comprising an open reading frame and a promoter and terminator non-covalently linked to said open reading frame.
Dependencies are inferred from the claim text.