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Vascular endothelial growth factor 2

A technology of nucleic acid molecules and myocardial ischemia, applied in the fields of epidermal growth factor, growth factor/inducer, cardiovascular system diseases, etc.

Inactive Publication Date: 2003-05-07
HUMAN GENOME SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although they are similar, VEGF and PDGF have specific differences

Method used

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  • Vascular endothelial growth factor 2
  • Vascular endothelial growth factor 2
  • Vascular endothelial growth factor 2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0413] Expression patterns of VEGF-2 in human tissues and breast cancer cell lines

[0414] Northern blot analysis was used to detect the expression level of VEGF-2 in human tissues and human breast cancer cells. RNAzol TMTotal RNA samples from cells were isolated by System B (Biotecx Laboratories, Ltd.). Use 1% agarose gel to separate the total amount of about 10 mg of RNA isolated from each breast cancer tissue and specific cell line, and spot it on a nylon filter, [Sambrook et al., Molecular Cloning: A Laboratory Manual, p. Second Edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York (1989)]. Labeling reactions were performed with 50 ng of DNA fragments according to the Stratagene Prime-It kit. Labeled DNA was purified from 5Prime-3Prime using Select-G-50 columns, Ltd. [Boulder, CO]. The filter was hybridized with the full-length VEGF-2 gene labeled with a radioactivity of 1,000,000 cpm / ml in 0.5M NaPO4 and 7% SDS solution overnight at 65°C. Wash twice a...

Embodiment 2

[0417] Expression of a truncated form of VEGF-2 (SED ID NO: 4) by in vitro transcription and translation

[0418] VEGF-2 cDNA was transcribed and translated in vitro to determine the size of the truncated form of VEGF-2 and the translatable polypeptide partially encoded by VEGF-2 cDNA. Two VEGF-2 inserts in the pBluescript SK vector were PCR amplified with three pairs of primers, 1) M13 reverse and forward primers; 2) M13 reverse primer and VEGF primer F4; and 3) M13 reverse primer and VEGF primer F5. These primer sequences are shown below.

[0419] M13 reverse primer: 5'-ATGCTTCCGGCTCGTATG-3' (SEQ ID NO: 9). This sequence is positioned upstream of the 5' end of the VEGF-2 cDNA insert in the pBluescript vector, as the cDNA it is in the antisense start orientation. A T3 promoter sequence was positioned between the primer and the VEGF-2 cDNA. M13-2 forward primer: 5'GGGTTTTTCCCAGTCACGAC-3' (SEQ ID NO: 10). This sequence was positioned downstream of the 3' end of the VEGF-2 ...

Embodiment 3

[0424] Cloning and expression of VEGF-2 using baculovirus expression system

[0425] With reference to ATCC No.97149, use the PCR oligonucleotide primer corresponding to this gene 5' and 3' sequence to amplify the DNA sequence of the VEGF-2c albumen of 46 amino acids without N terminal:

[0426] The 5' primer sequence is TGT AAT ACG ACT CAC TAT AGG GAT CCCGCC ATG GAG GCC ACG GCT TAT GC (SEQ ID NO: 12) and contains a BamH1 endonuclease site (in bold) and VEGF-25' Sequence complementary 17 nucleotide sequence (nt.150-166).

[0427] The 3' primer has the sequence GATC TCT AGA TTA GCT CAT TTG TGGTCT (SEQ ID NO: 13) and contains a restriction enzyme cleavage site XbaI and 18 nucleotides complementary to the VEGF-23' sequence including a stop codon and a stop codon The 15nt sequence.

[0428] Amplified fragments were separated from 1% agarose gels using a commercially available kit ("Geneclean," BIO101, Inc., La Jolla, CA). This fragment was digested with endonucleases BamH1 and ...

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Abstract

The invention discloses human VEGF-2 polypeptide, its biologically active fragments, analogues or derivatives which can be used for diagnosis or treatment, and DNA (RNA) encoding the VEGF-2 polypeptide. The invention also discloses the method for producing the polypeptide by recombinant technology, and the antibody and antagonist for the polypeptide. Such polypeptides and polynucleotides are useful therapeutically for stimulating wound healing and for the repair of vascular tissue. The present invention also discloses the use of the antibodies and antagonists to inhibit tumor angiogenesis and thereby inhibit tumor growth, inflammation, diabetic retinopathy, rheumatoid arthritis and psoriasis.

Description

Background of the invention [0001] The present invention relates to newly identified polynucleotides, polypeptides encoding the polynucleotides, uses of these polynucleotides and polypeptides, and production of these polynucleotides and polypeptides. The polypeptides of the present invention have been identified as members of the vascular endothelial growth factor family. More specifically, the polypeptide of the invention is human vascular endothelial factor 2 (VEGF-2). The invention also relates to the inhibition of the action of these polypeptides. [0002] The formation of new blood vessels, or angiogenesis, is a fundamental point in embryonic development, subsequent growth, and tissue repair. Angiogenesis is also an essential part of certain pathological conditions, such as neoplasia (ie, tumors and gliomas). Aberrant angiogenesis is also associated with other diseases such as inflammation, rheumatoid arthritis, psoriasis, and diabetic retinopathy (Folkman, J and Klags...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K35/76A61K38/00A61K45/00A61K48/00A61P9/00A61P9/06A61P9/10A61P17/06A61P27/02A61P29/00A61P31/04A61P31/10A61P31/12A61P33/00A61P35/00A61P37/06A61P37/08C07K14/485C07K14/52C12N1/15C12N1/19C12N1/21C12N5/10
CPCA01K2217/075A01K2217/05C07K14/52A61K48/00A61P9/00A61P9/06A61P9/10A61P17/06A61P27/02A61P29/00A61P31/04A61P31/10A61P31/12A61P33/00A61P35/00A61P37/06A61P37/08A61P43/00C12N15/11
Inventor 蒂莫西·A·科曼
Owner HUMAN GENOME SCI INC
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