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Polypeptide-human SEH protein 29 and polynucleotide encoding this polypeptide

A polynucleotide and protein technology, applied in the field of polynucleotide and polypeptide preparation, can solve the problem of low cyclooxygenase activity

Inactive Publication Date: 2004-01-28
BIOWINDOW GENE DEV INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Buehler et al. (1990) found that cyclooxygenase activity in amniotic cells of mothers taking phenytoin was low

Method used

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  • Polypeptide-human SEH protein 29 and polynucleotide encoding this polypeptide
  • Polypeptide-human SEH protein 29 and polynucleotide encoding this polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1: Cloning of human sEH protein 29

[0087] Total RNA was extracted from human fetal brain by one-step method of guanidine isothiocyanate / phenol / chloroform. Poly(A) mRNA was isolated from total RNA using Quik mRNA I isolation Kit (product of Qiegene). 2ug poly(A) mRNA was reverse transcribed to form cDNA. Smart cDNA cloning kit (purchased from Clontech) was used to insert the cDNA fragment into the multiple cloning site of the pBSK(+) vector (product of Clontech Company), transform DH5α, and the bacteria formed a cDNA library. The sequences of the 5' and 3' ends of all clones were determined using Dyeterminate cycle reaction sequencing kit (product of Perkin-Elmer) and ABI 377 automatic sequencer (Perkin-Elmer). Comparing the determined cDNA sequence with the existing public DNA sequence database (Genebank), it was found that the cDNA sequence of one of the clones, 0871b06, was a new DNA. The insert cDNA fragment contained in this clone was determined bidirec...

Embodiment 2

[0088] Example 2: Homology retrieval of cDNA clones

[0089] The sequence of the human sEH protein 29 of the present invention and the protein sequence encoded by it were published in Genbank, Databases such as Swissport conduct homologous searches. The gene with the highest homology to human sEH protein 29 of the present invention is a known human sEH protein, and the accession number of the encoded protein in Genbank is L05779. The results of protein homology are shown in Figure 1. The two are highly homologous, with an identity of 40%. Example 3: Cloning of the gene encoding human sEH protein 29 by RT-PCR

Embodiment 3

[0089] The sequence of the human sEH protein 29 of the present invention and the protein sequence encoded by it were published in Genbank, Databases such as Swissport conduct homologous searches. The gene with the highest homology to human sEH protein 29 of the present invention is a known human sEH protein, and the accession number of the encoded protein in Genbank is L05779. The results of protein homology are shown in Figure 1. The two are highly homologous, with an identity of 40%. Example 3: Cloning of the gene encoding human sEH protein 29 by RT-PCR

[0090] The total RNA of fetal brain cells was used as a template, and oligo-dT was used as a primer to carry out reverse transcription reaction to synthesize cDNA. After purification with a Qiagene kit, PCR amplification was performed with the following primers:

[0091] Primer1: 5'-GGCTCTTCTGGTCCCTGGTCTACTG-3'(3)

[0092] Primer2: 5'-TAATATAACTTTTTAATGGTTAAAA-3'(4)

[0093] Primer1 is the forward sequence starting from...

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Abstract

The present invention discloses a polypeptide-human sEH protein 29, polynucleotide for coding said polypeptide and method for producing said polypeptide by using DNA recombination technique. Said invention also discloses the method for using said polypeptide to curing several diseases, such as malignant tumor, hemopathy, HIV infection, immunological diseases and various inflammations. Said invention also discloses the agonist for resisting said polypeptide and its therapeutic action, and also discloses the application of polynucleotide coding said novel human sEH protein 29.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, the invention describes a polypeptide-human sEH protein 29, and a polynucleotide sequence encoding the polypeptide. The present invention also relates to the preparation method and application of the polynucleotide and polypeptide. Background technique [0002] Epoxygenases EH (EC 3.3.2.3) are a family of enzymes that hydrolyze various endogenous and exogenous epoxides to the corresponding diols. [0003] Cyclooxygenase exists widely in organisms, and cyclooxygenase can be found in various tissues of all animals that have been detected, and wherein the highest content is in liver and kidney [Wixtrom and HammockPharmacology and Toxicology (Zakim, D. and Vessey , D.A., ed.) 1:1-93, Wiley, New York, 1985]. Four major cyclooxygenases are known: leukotriene cyclooxygenase, cholesterol cyclooxygenase, microsomal cyclooxygenase and soluble cyclooxygenase (sEH). Amon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/00C07K14/435C07K14/47C07K16/18C12N15/11C12N15/113C12N15/55C12N15/63C12N15/64C12P21/02C12Q1/68
Inventor 毛裕民谢毅
Owner BIOWINDOW GENE DEV INC