Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
1999-02-05
2004-03-02
Eyler, Yvonne (Department: 1646)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S320100, C435S325000, C536S023500
Reexamination Certificate
active
06699660
ABSTRACT:
BACKGROUND
1. Technical Field
The present invention generally relates to gene expression and more specifically to immediate early genes in the brain and polypeptides encoded by such immediate early genes.
2. Background Information
An immediate early gene (IEG) is a gene whose expression is rapidly increased immediately following a stimulus. For example, genes expressed by neurons that exhibit a rapid increase in expression immediately following neuronal stimulation are neuronal IEGs. Such neuronal IEGs have been found to encode a wide variety of polypeptides including transcription factors, cytoskeletal polypeptides, growth factors, and metabolic enzymes as well as polypeptides involved in signal transduction. The identification of neuronal IEGs and the polypeptides they encode provides important information about the function of neurons in, for example, learning, memory, synaptic transmission, tolerance, and neuronal plasticity.
SUMMARY
The present invention involves methods and materials related to IEGs. Specifically, the invention provides isolated IEG nucleic acid sequences, cells that contain isolated IEG nucleic acid, substantially pure polypeptides encoded by IEG nucleic acid, and antibodies having specific binding affinity for a polypeptide encoded by IEG nucleic acid. In addition, the invention provides cDNA libraries enriched for IEG cDNAs, isolated nucleic acid derived from such cDNA libraries, and methods for treating conditions related to a deficiency in a neuron's IEG responsiveness to a stimulus.
In one aspect, the invention features an isolated nucleic acid having at least one adenine base, at least one guanine base, at least one cytosine base, and at least one thymine or uracil base. The isolated nucleic acid is at least 12 bases in length, and hybridizes to the sense or antisense strand of a second nucleic acid under hybridization conditions. The second nucleic acid has a sequence as set forth in SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60. The hybridization conditions can be moderately or highly stringent hybridization conditions.
In another embodiment, the invention features an isolated nucleic acid having a nucleic acid sequence that encodes an amino acid sequence at least five amino acids in length. The amino acid sequence contains at least three different amino acid residues, and is identical to a contiguous portion of sequence set forth in SEQ ID NO:11, 21, 27, 30, 32, 36, 38, 48, 61, or 62.
Another embodiment of the invention features an isolated nucleic acid having a nucleic acid sequence at least 60 percent identical to the sequence set forth in SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.
Another embodiment of the invention features an isolated nucleic acid having a nucleic acid sequence that encodes an amino acid sequence at least 60 percent identical to the sequence set forth in SEQ ID NO:11, 21, 27, 30, 32, 36, 38, 48, 61, or 62.
Another embodiment of the invention features an isolated nucleic acid having a nucleic acid sequence as set forth in SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.
In another aspect, the invention features a substantially pure polypeptide having an amino acid sequence encoded by a nucleic acid having at least one adenine base, at least one guanine base, at least one cytosine base, and at least one thymine or uracil base. The nucleic acid is at least 12 bases in length, and hybridizes to the sense or antisense strand of a second nucleic acid under hybridization conditions. The second nucleic acid has a sequence as set forth in SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.
In another embodiment, the invention features a substantially pure polypeptide having an amino acid sequence as set forth in SEQ ID NO:11, 21, 27, 30, 32, 36, 38, 48, 61, or 62.
Another embodiment of the invention features a substantially pure polypeptide having an amino acid sequence at least 60 percent identical to the sequence set forth in SEQ ID NO:11, 21, 27, 30, 32, 36, 38, 48, 61, or 62.
Another embodiment of the invention features a substantially pure polypeptide having an amino acid sequence at least five amino acids in length. The amino acid sequence contains at least three different amino acid residues, and is identical to a contiguous stretch of sequence set forth in SEQ ID NO:11, 21, 27, 30, 32, 36, 38, 48, 61, or 62.
Another aspect of the invention features a host cell (e.g., a eukaryotic or prokaryotic cell) containing an isolated nucleic acid having at least one adenine base, at least one guanine base, at least one cytosine base, and at least one thymine or uracil base. The isolated nucleic acid is at least 12 bases in length, and hybridizes to the sense or antisense strand of a second nucleic acid under hybridization conditions. The second nucleic acid has a sequence as set forth in SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.
Another aspect of the invention features an antibody (e.g., a monoclonal or polyclonal antibody) having specific binding affinity for an amino acid sequence encoded by a nucleic acid having at least one adenine base, at least one guanine base, at least one cytosine base, and at least one thymine or uracil base. The nucleic acid is at least 12 bases in length, and hybridizes to the sense or antisense strand of a second nucleic acid under hybridization conditions. The second nucleic acid has a sequence as set forth in SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 28, 29, 31, 33, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60.
Another aspect of the invention features a cDNA library having a plurality of clones with each clone having a cDNA insert. In addition, at least about 15 percent (e.g., at least about 20 or 25 percent) of the clones have cDNA derived from immediate early genes (e.g., immediate early genes responsive to a maximal electroconvulsive seizure). The cDNA library can be a subtracted cDNA library. For example, the subtracted cDNA library can be the IEG-Reg or IEG-Lg cDNA library.
Another aspect of the invention features an isolated nucleic acid derived from a cDNA library. The cDNA library has a plurality of clones with each clone having a cDNA insert. In addition, at least about 15 percent of the clones have cDNA derived from immediate early genes. The isolated nucleic acid can have a nucleic acid sequence of an immediate early gene.
Another aspect of the invention features a method of obtaining immediate early gene nucleic acid. The method includes providing a cDNA library having a plurality of clones with each clone having a cDNA insert. In addition, at least about 15 percent of the clones have cDNA derived from immediate early genes. The method also includes contacting at least a portion of the cDNA library with a probe containing at least one nucleic acid having a nucleic acid sequence derived from an immediate early gene, and selecting a member of the plurality of clones based on the hybridization of the at least one nucleic acid to the member under hybridization conditions.
Another aspect of the invention features a method of treating an animal (e.g., human) having a deficiency in a neuron's immediate early gene responsiveness to a stimulus. The method includes administ
Goetz Bernhard
Hiemisch Holger
Kuner Rohini
Lanahan Anthony
Nikolich Karoly
Eyler Yvonne
Gray Cary Ware & Freidenrich LLP
Haile Lisa A.
Murphy Joseph F.
Reynolds Kelly K.
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