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0072] Another method which may be used to retrieve unknown sequences is that of Parker, J. D. et al. (1991; Nuc. Acids Res. 19:3055-3060). Additionally, one may use PCR, nested primers, and PROMOTERFINDER libraries to walk in genomic DNA (Clontech, Palo Alto, Calif.). This process avoids the need to screen libraries and is useful in finding intron / exon junctions.
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For example, inappropriate cell survival can cause or contribute to many diseases such as cancer, autoimmune diseases, and inflammatory diseases.
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[0170] I HUVENOB01 cDNA Library Construction
[0171] Incyte clone 036596 was identified in the HUVENOB01 cDNA library prepared from control (or untreated) HUVEC cells. The HUVEC cell line is a normal, homogeneous, well-characterized early passage endothelial cell culture from human umbilical vein (Cell Systems Corp., Kirkland, Wash.).
[0172] The induced human endothelial cell (HUVEC) cDNA library was custom constructed by Stratagene (Cat. #937207). cDNA synthesis was primed using either oligo d(T) or random hexamers. Synthetic adaptor oligonucleotides were ligated onto cDNA ends enabling insertion into the lambda vector. The HUVEC cDNA library was constructed using the UNIZAP vector system (Stratagene). Finally, the two libraries were combined into a single library by mixing equal numbers of bacteriophage.
[0173] The custom-constructed library phage particles were infected into E. coli host strain XL1-BLUE (Stratagene) which has a high transformation efficiency, increasing the probabili...
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Abstract
The present invention provides a human apoptosis-related calcium-binding protein (HARC) and polynucleotides which identify and encode HARC. The invention also provides genetically engineered expression vectors and host cells comprising the nucleic acid sequences encoding HARC and a method for producing HARC. The invention also provides for agonists, antibodies, or antagonists specifically binding HARC, and their use, in the prevention and treatment of diseases associated with expression of HARC. Additionally, the invention provides for the use of antisense molecules to polynucleotides encoding HARC for the treatment of diseases associated with the expression of HARC. The invention also provides diagnostic assays which utilize the polynucleotide, or fragments or the complement thereof, and antibodies specifically binding HARC.
Description
[0001] This application is a continuation application of U.S. Ser. No. 09 / 328,687, filed Jun. 9, 1999, which is a divisional application of U.S. Ser. No. 09 / 094,212, filed Jun. 9, 1998, now U.S. Pat. No. 5,935,931, which is a divisional application of U.S. Ser. No. 08 / 766,605, filed Dec. 12, 1996, now U.S. Pat. No. 5,763,220, the contents each of which is hereby incorporated by reference, and each application entitled "Human Apoptosis-Related Calcium-Binding Protein".[0002] This invention relates to nucleic acid and amino acid sequences of a novel human apoptosis-related calcium-binding protein and to the use of these sequences in the diagnosis, prevention, and treatment of diseases associated with decreased or increased apoptosis.[0003] Normal development, growth, and homeostasis in multicellular organisms require a careful balance between the production and destruction of cells in tissues throughout the body. Cell division is a carefully coordinated process with numerous checkpoin...
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Patent Type & Authority Applications(United States)
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