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Gene order of b subtype of 4 receptor of hydroxytryptarmine in pig

A technology of gene sequence and serotonin, applied in the field of protein coding sequence, can solve the problem of undiscovered clone coding protein sequence, etc.

Inactive Publication Date: 2005-01-12
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In the search and analysis of prior art documents, although "FEBS Letters, 1995, 370: 215-221" (Expression of serotonin receptor mRNAs in blood vessels, the expression of serotonin receptor mRNAs in blood vessels) by RT-PCR The partial coding protein sequence of porcine 5-serotonin 4 receptor was cloned by the method, and the expression of 5-serotonin 4 receptor gene in human, mouse, pig and other species was analyzed, but so far no porcine 5-serotonin has been cloned The Complete Coding Protein Sequence of the 4 Receptor Gene and the Report on the Evolution of Mammalian 5HT 4 Receptor Gene

Method used

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  • Gene order of b subtype of 4 receptor of hydroxytryptarmine in pig
  • Gene order of b subtype of 4 receptor of hydroxytryptarmine in pig
  • Gene order of b subtype of 4 receptor of hydroxytryptarmine in pig

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Embodiment 1

[0125] Cloning of cDNA Sequence of Porcine 5-HT4 Receptor Subtype b Gene

[0126] 1. Isolation

[0127] The 30-day-old Dudu piglets were slaughtered in the pig farm, and the colon tissue samples were taken, quickly put into liquid nitrogen, frozen and stored in the -70°C refrigerator in the laboratory, ready to extract total RNA from the tissue.

[0128] 2. Total RNA isolation (Total RNA isolation)

[0129] (1) Preparation for RNA extraction

[0130] Glass products were soaked in 0.1M NaOH overnight, rinsed repeatedly with tap water, rinsed twice with distilled water, and baked at 180°C for 4 hours.

[0131]Soak the homogenizer and electrophoresis tank with 3% hydrogen peroxide for 20-30 minutes, and then rinse with 0.1% DEPC water. Since the uninactivated DEPC will affect the PCR reaction, it can be treated with 0.5% SDS.

[0132] Soak the Tip and EP tube in 0.1% DEPC water overnight, and sterilize by autoclaving to inactivate DEPC.

[0133] The double-distilled water and...

Embodiment 2

[0151] Sequence Analysis and Homology Comparison of Porcine 5-HT4 Receptor Subtype b Gene

[0152] 1. Gene sequence analysis of porcine serotonin 4 receptor b subtype

[0153] The length of the cDNA of the novel porcine serotonin 4 receptor b subtype gene of the present invention is 1029bp, and the detailed sequence is shown in SEQ ID NO.7, wherein the open reading frame is located at 1-1029 nucleotides (1029 nucleotides). The amino acid sequence of porcine serotonin 4 receptor b subtype was deduced according to cDNA, with a total of 402 amino acid residues, a molecular weight of 44960 Daltons, and an isoelectric point (pI) of 8.08. See SEQ ID NO.7 for the detailed sequence.

[0154] 2. Homology comparison of serotonin 4 receptor b subtype gene

[0155] The cDNA sequence of porcine serotonin 4 receptor b subtype gene and its encoded protein were analyzed by BLAST program in Non-redundant GenBank +EMBL+DDBJ+PDB and Non-redundant GenBank CDStranslations +PDB+ SwissProt+Superda...

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Abstract

This invention relates to a serotonin 4 receptor b subtype gene sequence of pigs amplifying a serotonin 4 receptor b subtype gene nucleotide sequence form a master RNA of a pig's connective tissue, the tolal length of which is 1209bp, the open reading frame 1bp-1209bp segments coded with 402 amino acid protein, the molecular weight is 44960 dalton, pl is 8.08, which is important to the regulation of stomach intestine movement of pigs and an important standby gene for studying pig stress diarrhea and pig molecular marked assistant breeding.

Description

technical field [0001] The invention relates to a protein coding sequence used in the technical field of genetic engineering, in particular to a gene sequence of pig serotonin 4 receptor b subtype. Background technique [0002] No matter in human medicine or animal medicine, stress diarrhea is a common clinical disease, and diarrhea in weaned piglets is a typical example of animal stress diarrhea. Although the understanding and research on stress diarrhea has gone through more than a century, its etiology and pathogenesis are still unclear. In recent years, some scholars at home and abroad have explored the biological effects of neurotransmitter 5-HT and 5-HT-4 receptors in mediating stress diarrhea. [0003] 5-hydroxytryptamine receptor 4 (5-hydroxytryptamine receptor 4, 5-HT4R) is a class of binding receptors for the neurotransmitter 5-hydroxytryptamine, which belongs to the G protein-coupled receptor superfamily and is widely distributed in the central nervous system and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12
Inventor 潘玉春赵威孟和白春艳李婧
Owner SHANGHAI JIAO TONG UNIV