N-end fusion protein PC-1-e2 of human multicapsular protein-1

A polycystin and fusion protein technology, applied in the field of medical molecular bioengineering, can solve the problems of high cost, short half-life, and limited intervention effect, and achieve low cost and long-lasting inhibitory effect

Inactive Publication Date: 2004-09-08
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology

[0004] Intervention measures to inhibit abnormal proliferation of renal tubular epithelial cells have been reported, such as lovastatin, tyrosine protein kinase inhibitors, etc., which are not only expen

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  • N-end fusion protein PC-1-e2 of human multicapsular protein-1

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Embodiment Construction

[0025] The preparation of human PC-1 N-terminal fusion protein PC-1-e2 is now described in detail.

[0026] 1. Materials

[0027] 1. Plasmids and strains: fusion protein expression vector pQE30 (the 5' end of its reading frame contains a nucleotide sequence encoding 6 consecutive histidines, and the plasmid contains an ampicillin resistance marker gene) and expression host bacterium M15 (the strain A repressor plasmid pREP4, which contains a kanamycin resistance marker gene) was purchased from Qiagen, Germany.

[0028] 2. Main reagents and instruments The total RNA extraction kit and gel recovery kit are products of Huashun Company, and the One-Step RT-PCR kit and Ni-NTA purification reagent (Ni-NTA Agarose) were purchased from Qiagen, Germany. Various restriction enzymes, T 4 DNA ligase is a product of Takara Company. Ampicillin (Amp) and Kanamycin (Kan) were purchased from Shanghai Huamei Bioengineering Company. Fetal bovine serum, RPMI 1640 and DMEM cell culture medium ...

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Abstract

The present invention relates to medicine molecular biology engineering technology, and is one kind of 1N-end fusion protein PC-1-e2 of human polycystic protein-1. According to cDNA sequence of human polycystic kidney disease-1 gene and the molecular structure of its encoded product, polycystic protein-1 (PC-1), the gene is cloned to cDNA-PKD1e2 in the N end of the protein and its prokaryotic expression carrier pQE30-PKD1e2 is constituted by means of molecular biology engineering technology. N-end fusion protein PC-1-e2 of PC-1 is induced in colibacillus. Cytobiology experiment shows that the fusion protein has obvious inhibition to the proliferation of the epidermis cell line of in vitro cultured human polycystic kidney cyst lining and the epidermis cell strain of far end renal tubule in dog kidney, so that it may be used in preparing medicine for treating autosomal dominant hereditary polycystic kidney disease, the reagent for the mechanism research of the disease, and anti-PC-1 antibody.

Description

technical field [0001] The invention relates to the technical field of medical molecular bioengineering, which is a human polycystin-1 N-terminal fusion protein PC-1-e2, which can be used to prepare medicine for treating autosomal dominant polycystic kidney disease, and can be used to prepare and study the Reagents of disease pathogenesis and preparation of anti-PC-1 antibodies. Background technique [0002] Autosomal dominant polycystic kidney disease (ADPKD) is a hereditary kidney disease that seriously endangers human health. Patients with bilateral kidneys are densely covered with cysts, and often develop into end-stage renal failure in late middle age. Due to its The pathogenic gene is widely expressed in the body, and often involves other epithelial organs, including the liver, pancreas, ovary and choroid plexus, etc., causing great harm. It is now clear that 85-90% of the occurrence of ADPKD is due to the structural and functional abnormal...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P13/12C07K16/18C07K19/00G01N33/68
Inventor 赵海丹梅长林
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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