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Chicken GBP4L gene, expression vector and application

A technology for expressing vectors and genes, applied in the field of genetic engineering

Active Publication Date: 2022-08-09
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] How to alleviate the heat stress of broilers has always been a difficult problem in the breeding industry. Traditional ways to alleviate heat stress, such as increasing the energy concentration of the diet, or applying additives such as electrolytes, minerals, vitamins, and Chinese herbal medicine preparations, have limitations in terms of application and function. some limiting factors

Method used

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  • Chicken GBP4L gene, expression vector and application
  • Chicken GBP4L gene, expression vector and application
  • Chicken GBP4L gene, expression vector and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Chicken GBP4L gene

[0040] The nucleotide sequence of the chicken GBP4L gene of the present embodiment is shown in SEQ ID NO: 1, and the nucleotide sequence and the corresponding amino acid sequence of the chicken GBP4L gene are as follows: figure 1 shown.

Embodiment 2

[0041] Example 2 Expression vector containing chicken GBP4L gene

[0042] The expression vector containing the chicken GBP4L gene in this example is obtained by connecting the chicken GBP4L gene to an adeno-associated virus vector (AAV). The vector maps of the tool vector and the final vector used in this example are respectively as follows figure 2 , image 3 shown.

[0043] The GBP4L sequence was synthesized by Shanghai Shenggong and constructed into the target vector. Screen the positive clones and verify them by sequencing. The sequencing results of the positive clones are as follows: Figure 4 shown (SEQ ID NO: 2), indicating that the pAAV-GBP4L-3FLAG adeno-associated virus recombinant plasmid was successfully constructed.

[0044] 1. Recombinant AAV virus release and packaging

[0045] Use HEK293 cell line to produce adeno-associated recombinant virus, HEK293 cell is a cell line obtained by transforming human embryonic kidney cells with sheared adenovirus type 5 DN...

Embodiment 3

[0084] Example 3 Application in Relieving Chicken Heat Stress and Breeding of Chicken Breeds

[0085] 3.1. Animal experiment design and sample preparation

[0086] Forty-eight healthy Gushi roosters with similar body weight at 28 days were selected as the research objects, placed in an isolated clean chicken coop, and raised in cages, and the light, temperature and humidity were strictly controlled during the raising process. They were randomly divided into four groups, A, B, C, and D, with 12 chickens in each group. Group A was the control group, group B was the heat stress group, group C was the heat stress + control AAV group (no GBP4L recombinant fragment AAV, pAAV-3FLAG), and group D was the heat stress + overexpressing GBP4L recombinant AAV Group (insert GBP4L recombinant fragment AAV, pAAV-GBP4L-3FLAG). Group C was given subcutaneous injection of purified pAAV-3FLAG virus in the neck, and the virus amount was 2.43 * 10 10 vg; Group D was given subcutaneous injection...

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Abstract

The invention belongs to the technical field of gene engineering, and particularly relates to a chicken GBP4L gene, an expression vector and application. The nucleotide sequence of the chicken GBP4L gene is as shown in SEQ ID NO: 1. It is proved for the first time that the GBP4L gene is related to chicken heat stress, and phenotypes such as inflammatory factor increase and immune organ injury caused by chicken heat stress can be effectively relieved by regulating and controlling high expression in vivo of the GBP4L gene. The research result of the invention has a good application prospect in the aspects of chicken heat stress relieving and heat-stress-resistant chicken variety breeding.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a chicken GBP4L gene, an expression vector and an application. Background technique [0002] Heat stress seriously affects the development of animal husbandry and causes major losses to the global breeding industry every year. The current mainstream livestock and poultry breeds cannot effectively dissipate heat due to the lack of sweat glands on their skin surfaces, and are sensitive to changes in ambient temperature. It has adverse effects on livestock and poultry production. Under heat stress, the body's electrolyte balance, glucose and lipid metabolism, antioxidant system and intestinal health are all negatively affected, which in turn leads to low mood, loss of appetite, and reduced immunity in livestock and poultry. [0003] How to alleviate the heat stress of broilers has always been a difficult problem in the breeding industry. The traditional ways...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/864C12N7/01A01K67/027C12R1/93
CPCC07K14/465C12N15/86C12N7/00A01K67/0278C12N2750/14121C12N2750/14143A01K2227/30A01K2267/02Y02P60/87
Inventor 康相涛郭玉洁田亚东闫峰宾李国喜蒋瑞瑞韩瑞丽孙桂荣李文婷李东华张彦华李转见李红刘小军
Owner HENAN AGRICULTURAL UNIVERSITY