Inducer for activating expression of fish lipid metabolism control gene and application thereof

A technology for regulating genes and inducers, applied in the field of biotechnology applications, can solve the problems of slow growth, abnormal body shape, and reduce the quality and flavor of fish meat, and achieve the effects of improving meat quality and reducing fat deposition.

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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005]Grass carp (Ctenopharyngodon idella) is the main breeding target in my country, and it can effectively use cheap plant protein sources to provide aquatic products. Increased body fat content and the occurrence of fatty liver are more prominent
Under intensive farming conditions, due to a large intake of artificial compound feed and lack of exercise, grass carp body fat deposits, slow growth, and abnormal body shape, which reduces the quality and flavor of fish meat, and also increases the cost of farming

Method used

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  • Inducer for activating expression of fish lipid metabolism control gene and application thereof
  • Inducer for activating expression of fish lipid metabolism control gene and application thereof
  • Inducer for activating expression of fish lipid metabolism control gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 The successful cloning of the new cDNA sequence of grass carp peroxisome proliferator-activated receptor gene

[0044] Grass carp was provided by Guangdong Famous Freshwater Fish Breeding Center (Panyu, Guangdong).

[0045] (1) Total RNA extraction and first-strand cDNA synthesis

[0046] Liver tissue was quickly isolated from grass carp, and total RNA was extracted and purified according to the method recommended by Promega's SV Total RNA Isolation System kit. The first strand of cDNA was synthesized using the ReverTra Ace-α-TM kit from TOYOBO, with grass carp liver total RNA as the template and oligo (dT)20 as the reverse transcription primer, and the operation was carried out according to the method recommended by the kit.

[0047] (2) Cloning of the cDNA core sequence of grass carp PPAR gene

[0048] Three pairs of degenerate primers PPARα01F and PPARα02R, PPARβ01F and PPARβ02R, PPARγ01F and PPARγ02R were designed and synthesized based on the conserved r...

Embodiment 2

[0054] Example 2 Study on tissue expression of grass carp PPARα, PPARβ and PPARγ genes

[0055] Using β-actin as an external reference, the relative expression levels of PPARα, PPARβ and PPARγ gene mRNA in grass carp liver, brain, spleen, muscle, fat and heart were compared by fluorescent quantitative PCR method, with 5 records in each group. Specific primers FQPPARα01F and FQPPARα02R, FQPPARβ01F and FQPPARβ02R, FQPPARγ01F and FQPPARγ02R were designed according to the cDNA core sequences of grass carp PPARα, PPARβ and PPARγ genes (Table 1), and the PCR product fragment sizes were 202 bp, 185 bp and 206 bp, respectively. Specific primers FQACT01F and FQACT02R were designed for β-actin (Table 1). The PCR reaction conditions were 94 °C pre-denaturation for 3 min, 94 °C for 1 min, 57 °C for 1 min, 72 °C for 1 min, a total of 30 cycles, and the final cycle at 72 °C Extend for 5 min. For the PPAR / beta-actin mRNA (%) data of grass carp liver, brain, spleen, muscle, fat and heart, ple...

Embodiment 3

[0056] Example 3 The effect of intraperitoneal injection of different concentrations of inducers on the expression of PPAR gene in grass carp

[0057] Grass carp are raised in aquariums with constant flow of filtered water, and rationed daily from 10:00 to 11:00 with floating pellets of 2% of the fish’s body weight (Guangzhou Haiwei), collected at 11:00-12:00 Remaining feed, 7 days before the experiment, re-weighed, and grouped fish with similar body weight into one group, with 5 fish in each group, for the injection test. The inducer Clofibrate (clofibrate) was dissolved in dimethylsulfoxide, 2-bromo palmitate (2-bromopalmitate), 15-deoxy-Δ12,14-prostaglandin J2 (15-deoxy-Δ12,14-prostaglandin Primer J2) was dissolved in absolute ethanol. Phosphate buffer PBS was used as the diluent, and the inducer was diluted to the corresponding concentration. After feeding once at 10:00-11:00, the anesthesia injection was started. The control group was only injected with 200ulPBS, and th...

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Abstract

The invention discloses an inducer for activating the expression of a fish lipid metabolism control gene and application thereof. The inducer is prepared from one or a mixture of more of clofibrate, 2-palmitic acid, 15-deoxidized-delta 12 and 14-prostaglandin J2. The inducer can activate the expression of the fish somatic cell lipid metabolism control gene, namely a peroxisome proliferator activated receptor gene, so that the inducer can be used for breeding fishes, maintaining the nutrition metabolization balance in the fish breeding, reducing lipidosis of the fishes and improving the quality of fish flesh.

Description

technical field [0001] The invention relates to the application field of biotechnology, in particular to an inducer for activating fish lipid metabolism regulation gene expression and application thereof. Background technique [0002] Peroxisome proliferator activated receptors (PPARs) are a class of nuclear receptors that can be activated by endogenous fatty acids and exogenous peroxisome proliferators (peroxisome proliferators, PP) as ligands. Transcription factor, a member of the type II nuclear receptor superfamily. PPAR is also called fatty acid receptor because it can be activated by endogenous fatty acids and their metabolites. There are three types of PPARs receptors in various animals, namely PPARα, PPARβ, also known as PPARδ and PPARγ. These three subtypes are specific and can produce different biological effects. PPAR can regulate the expression of many target genes involved in intracellular and extracellular lipid metabolism, especially the genes encoding some ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/216A61K31/19A61K31/20A61P3/00C12N15/09A23K1/18A23K20/10A23K50/80
Inventor 梁旭方何珊陈小佳邹奕陈亮姚煜郁颖
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