New application of beta-N-acetylglucosaminyl transferase

A technology of glucose transferase and acetylaminotransferase is applied in the field of beta-N-acetylglucosamine transferase to achieve the effect of improving cold tolerance

Pending Publication Date: 2016-08-17
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The role of OGT in animal cold tolerance has not been reported

Method used

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  • New application of beta-N-acetylglucosaminyl transferase
  • New application of beta-N-acetylglucosaminyl transferase
  • New application of beta-N-acetylglucosaminyl transferase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] 1. Isolation and culture of mouse skeletal muscle satellite cells

[0017] Select Kunming mice aged 5-10 days, 75% alcohol immersion disinfection, skin peeling under aseptic conditions, complete removal of the hind limb muscles, back muscles, oblique shoulder muscles and diaphragm muscles of newborn mice, and divide them into small pieces Block, perform the first digestion (Ham's F-10 medium: 500U / ml collagenase II, 10% horse serum and 1% penicillin / streptomycin) and digestion in a 37℃, 5% CO2 cell incubator 90min; then add an equal volume of detergent (Ham's F-10 containing 10% horse serum) to terminate the digestion, and let it stand for 1 minute to make the large flakes (undigested fibers contain fibroblasts) fall into the bottom of the tube. The supernatant contains single fiber fragments, carefully transfer to a new 15ml centrifuge tube, 500g separation for 1min, discard the supernatant (the first wash), and then repeat the wash twice; for the second digestion (Hams F...

Embodiment 2

[0021] A total of 15 healthy male Kunming mice weighing (18±2)g at 6 weeks of age were selected, purchased from Jilin Changchun Animal Test Base, raised in artificial intelligence climate indoor, feed intake: 5g / 100g, free drinking water, temperature: low temperature ( 4±0.1)℃; normal temperature (21±0.1)℃, humidity (40±0.1)%. SD mice were randomly divided into: A normal temperature control group; B low temperature stress group for 4 hours, C low temperature stress group for 8 hours, 5 mice in each group. Groups B and C were reared at 4±0.05℃, and the cold stimulation time was 4h and 8h, respectively. Group A was kept in a room temperature environment, and the temperature was 21±0.05℃.

[0022] Fluorescence quantitative PCR to detect the expression of OGT and OGA:

[0023] In vitro experiments, the cells of each test group were collected at room temperature, cold stress for 4h and 8h respectively, and 1mL of Trizol reagent was added to the collected cells and placed at room tempe...

Embodiment 3

[0029] ELISA method was used to detect the concentration of Casp-3 in mouse skeletal muscle satellite cells of each group. The result is figure 1 As shown, under the normal incubation temperature of 37°C, compared with the normal control group, the Casp-3 concentration of the OGT high expression treatment group decreased, and the difference was significant (P Figure 5 Shown.

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Abstract

The invention relates to an application of beta-N-acetylglucosaminyl transferase to cold-resistant medicine preparing. According to the application, the OGT expression quantity and the time of cold stress are positively related, the OGT expression quantity after the cold stress is increased, it can be protected that skeleton satellite cells are not damaged by the cold stress, and the cold-resistant effect on skeletal muscle can be achieved; OGT is injected into the bodies of mice, and the cold-resistant capacity of the mice can be obviously improved; in the way, the OGT can serve as cold-resistant medicine to be popularized in a large-scale mode.

Description

Technical field [0001] The invention belongs to the field of animal molecular biology and relates to a new use of β-N-acetylglucosamine transferase. Background technique [0002] Cold stimulation is the most common stressor for animals in the northern cold regions. The breeding of livestock and poultry in winter, especially sows in early spring and late autumn, calves in winter, and chicks hatched in early spring, will cause cold stress in animals due to low temperatures. (Cold stress). Animal nutrition and metabolism disorders, slow growth, poor disease resistance and even death caused by cold stress are one of the main factors restricting the development of animal husbandry and breeding in the north. [0003] Skeletal muscle is the main metabolic tissue of the body, and plays a pivotal role in exercise, heat production and energy balance. A full understanding of the molecular mechanism of nutrient flux in muscle will play an important role in promoting muscle growth and improvin...

Claims

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Application Information

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IPC IPC(8): A61K38/45A61P39/00
CPCA61K38/45C12Y204/01
Inventor 李士泽张雪杨玉英甄莉计红杨焕民李鹏姚睿智刘洋
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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