Novel hemopoietin receptor protein, nr12

Abstract

A novel hemopoietin receptor gene (NR12) was successfully isolated by extracting motifs conserved among the amino acid sequences of known hemopoietin receptors and by using the predicted sequence. The NR12 gene encodes two forms of proteins, a transmembrane type and a soluble type. The expression of the NR12 gene was detected in tissues containing hematopoietic cells. NR12 is a novel hemopoietin receptor molecule involved in the regulation of immune system and hematopoiesis in vivo. Thus, NR12 is useful in the search for novel hematopoietic factors that functionally bind to the NR12 receptor, and in the development of therapeutic drugs for diseases associated with immunity or hematopoiesis.

Description

[0001]This application is a divisional of U.S. application Ser. Nos. 12 / 205,799 and 12 / 205,753, both filed Sep. 5, 2008, each being a divisional of U.S. application Ser. No. 11 / 595,320, filed Nov. 9, 2006, which is a continuation of U.S. application Ser. No. 11 / 274,375, filed Nov. 14, 2005, now abandoned, which is a divisional of U.S. application Ser. No. 10 / 105,930, filed Mar. 25, 2002, now U.S. Pat. No. 7,045,595, which is a continuation-in-part of International Patent Application No. PCT / JP00 / 06654, filed Sep. 27, 2000, which claims the benefit of Japanese Patent Application Nos. 11-273358, filed Sep. 27, 1999, and 2000-240397, filed Aug. 3, 2000. Each of the prior U.S. applications is herein incorporated by reference.TECHNICAL FIELD[0002]The present invention relates to novel hemopoietin receptor proteins, and genes encoding them, as well as methods for producing and using the same.BACKGROUND[0003]A large number of cytokines are known as humoral factors that regulate proliferati...

AI Technical Summary

Benefits of technology

[0038]Analyses of the gene expression in various human organs using RT-PCR revealed: strong expression of NR12 in hematopoietic cell line tissues and immune cell line tissues such as adult spleen, thymus, lymph node, bone marrow, and peripheral leukocyte; and expression in testis, liver, lung, kidney, pancreas, and gastrointestinal tract, such as small intestine and colon. Additionally, expression of NR12 was also observed in all the analyzed mRNA derived from human fetal organs. From the revealed distribution pattern of NR12 gene expression, it was presumed that NR12 encodes a novel hematopoietic factor receptor, primarily because localization of strong expression in tissues thought to include immune cell lines and hematopoietic cells was detected. Furthermore, the fact that NR12 expression was observed in tissues other than those described above suggests that NR12 can regulate not only physiological functions of the immune system and hematopoietic system in vivo but also various other physiological functions in vivo.

[0077]Moreover, the present invention provides a vector containing a DNA of the present invention as an insert. The vector of the present invention may be useful for maintaining the DNA of the present invention in host cells or producing the protein of the present invention.

[0131]An antisense oligonucleotide derivative is given to the patient by directly applying it onto the ailing site, by injecting it into the blood vessel and such, so that it will reach the ailing site. An antisense-mounting medium can also be used to increase durability and membrane-permeability. Examples are, liposome, poly-L-lysine, lipid, cholesterol, lipofectin or derivatives of them.

[0142]SDS-PAGE is generally used for the analysis of immunoprecipitated proteins, and bound proteins can be analyzed based on the molecular weight of proteins using a gel of an appropriate concentration. In this case, although proteins bound to a protein of the present invention, in general, are hardly detectable by the usual protein staining methods, such as Coomassie staining and silver staining, the detection sensitivity can be improved by culturing cells in a culture medium containing radio isotope-labeled 35S-methionine and 35S-cystein to label proteins inside the cells, and detecting the labeled proteins. Once the molecular weight of the protein is determined, the desired protein can be purified directly from SDS-polyacrylamide gel and sequenced.

Problems solved by technology

However, it was extremely difficult to strictly select only those to which all 15 nucleotides that encode the motif would completely hybridize under the usual hybridization conditions, because the oligonucleotide “tggag(t / c)nnntggag(t / c)” (wherein “n” is an arbitrary nucleotide; SEQ ID NO:22) encoding the motif was short, having just 15 base pairs.

Therefore, performing screening under normal hybridization experiment conditions was extremely difficult.

However, this YR motif is not exactly a perfect consensus sequence, and the combination of the nucleotide sequences that encode the motif is very complicated.

Therefore, it is practically impossible to synthesize and provide oligonucleotides that encode all of the amino acid sequences as probes for hybridization, which is a practical method for screening, or as primers aimed for RT-PCR.

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