Yao horsefly salivary gland immunoregulation peptide, gene and application thereof

A technology of immune regulation and salivary glands, which is applied in the field of biomedicine, can solve problems such as no public reports, and achieve the effects of strong antibacterial activity, simple structure, and convenient artificial synthesis

Active Publication Date: 2008-07-16
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Through document retrieval, do not see the public report identical with the present invention

Method used

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  • Yao horsefly salivary gland immunoregulation peptide, gene and application thereof
  • Yao horsefly salivary gland immunoregulation peptide, gene and application thereof
  • Yao horsefly salivary gland immunoregulation peptide, gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Cloning of Yaofly salivary gland immunomodulatory peptide gene:

[0032] I. Extraction of total RNA from the salivary glands of Yaofly:

[0033] A. Live dissecting Yaofly, pick salivary glands and put them into liquid nitrogen immediately, accumulate about 60, weigh, get 300mg Yaofly salivary gland tissue, add 10ml total RNA extraction buffer (Trizol solution, the product of American GIBCOBRL company), Homogenize for 30 minutes in a 20ml glass homogenizer.

[0034] B. Add an equal volume of phenol / chloroform solution, mix vigorously, leave at room temperature for 10 minutes, centrifuge at 12,000 rpm for 10 minutes at 4°C, and discard the precipitate.

[0035] C. Add an equal volume of isopropanol to the supernatant, leave it at room temperature for 10 minutes, centrifuge at 12,000 rpm at 4°C for 10 minutes, wash the precipitate once with 75% ethanol, and dry it in the air.

[0036] II. Purification of Yaofly salivary gland mRNA:

[0037] Yaofly salivary gl...

Embodiment 2

[0106] Example 2: Preparation of Yaofly Salivary Gland Immunomodulatory Peptide:

[0107] I. The preparation method of the Yaofly salivary gland immunomodulatory peptide: the amino acid sequence of the Yaofly salivary gland immunomodulatory peptide is deduced according to the gene encoding the Yaofly salivary gland immunomodulatory peptide, and its full sequence is synthesized with an automatic polypeptide synthesizer. Desalted and purified by HPLC reverse phase C18 column chromatography.

[0108] II. The molecular weight is determined by fast atom bombardment mass spectrometry (FAB-MS), with glycerol: m-nitrobenzyl alcohol: dimethyl sulfoxide (1: 1: 1, V: V: V, volume ratio) as substrate, Cs + As the bombardment particles, the current is 1μA and the emission voltage is 25Kv.

[0109] III. The purity of the purified Yaofly salivary gland immunomodulatory peptide was identified by high performance liquid chromatography (HPLC), the molecular weight was determined by fast atom ...

Embodiment 3

[0110] Example 3: Pharmacological experiment of Yaofly salivary gland immunomodulatory peptide:

[0111] 1. The inhibitory effect of Yaofly salivary gland immunomodulatory peptide on tumor necrosis factor (TNF-α) (Figure 1):

[0112] Preparation and culture of rat splenocytes: Rats were sacrificed by breaking the cervical spine, the spleen was aseptically removed and the adipose tissue was peeled off, the blood was washed away in RPMI1640 medium, the spleen was aseptically cut up and transferred to a 200-mesh copper mesh, and cored with a 2ml syringe Spleens were ground to disperse single cells. Centrifuge at 1000rpm / min for 10 minutes, discard the supernatant, suspend the pellet in RPMI1640 medium containing 10% fetal bovine serum and 2ug / ml bacterial lipopolysaccharide (LPS), adjust the cell concentration to 2×10 6 pieces / ml. Add 196ul of cell suspension to each well of the 96-well plate, and add 4ul of gadfly immunomodulatory peptide with a concentration of 1mg / ml to make...

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PUM

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Abstract

The invention relates to a salivary gland immunoregulation polypeptide of a dieu cleg and the related genes and the application, belonging to the biomedical field. The immunoregulation polypeptide is a linear polypeptide coded with the Chinese Tabanidae salivary gland immunoregulation polypeptide genes, the sequence of which is glycin- glycin- valine- serine- glycin- valine- serine- aspartic acid- phenylalanine- glutamate- praline- isoleucine- glutamate- valine- serine- glycin- glutamate- aspartic acid- tyrosine- aspartic acid- serine- aspartic acid- glutamate- alanine- aspartic acid- glutamate- aspartic acid- glycin- lysine- alanine. The immunoregulation polypeptide gene is composed of 362 Nucleotides, wherein, the coding mature code salivary gland immunoregulation polypeptide of the dieu cleg is the 115th to the 204th nucleotide. The dieu cleg salivary gland immunoregulation polypeptide is used for inhibiting a plurality of cytokines, which is characterized by simple structure, convenient artificial synthesis and strong antibacterial activity, etc.

Description

Technical field: [0001] The invention provides a Yaofly salivary gland immunomodulatory peptide and its gene and application, which belong to the technical field of biomedicine. Background technique: [0002] Immunomodulatory peptides have a wide range of biological activities at their physiological concentrations, in the presence of salt and serum, including in innate immune cells (neutrophils, epithelial cells) and inflammatory responses, innate immunity and adaptive Immune-related activity in monocytes, macrophages, and dendritic cells as a bridge between innate and adaptive immunity. Antimicrobial peptides can neutralize G - Lipopolysaccharide, G + Bacterial signaling molecules such as lipoteichoic acid and bacterial unmethylated CpG DNA. These signaling molecules can bind to Toll-like receptors on the surface of host cells to initiate signal cascade amplification and positive regulation of cytokines (such as TNF-α, IL-6, etc.). Low concentrations of bacterial signal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/10A61P37/02
Inventor 赖仞武静徐学清李东升马冬莹
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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