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Gene chip for evaluating transport stress model and application of gene chip for evaluating transport stress model

A transport stress and gene chip technology, which is applied in the field of gene chips for evaluating transport stress models, can solve the problems of unstable and precise indication of stress state, many influencing factors, and fast metabolism.

Inactive Publication Date: 2013-04-10
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no definite standard for the evaluation of the transport stress model. Most of the time, the changes in hormone levels such as cortisol and catecholamines are used to measure the occurrence of stress. However, there are too many factors that cause hormone changes, and their own metabolic rate fast, and cannot stably and accurately indicate the actual stress state of the body, so we introduce the new technology of gene chip

Method used

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  • Gene chip for evaluating transport stress model and application of gene chip for evaluating transport stress model
  • Gene chip for evaluating transport stress model and application of gene chip for evaluating transport stress model
  • Gene chip for evaluating transport stress model and application of gene chip for evaluating transport stress model

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Embodiment 1

[0056] The preparation of embodiment 1 gene chip

[0057] Regarding the preparation of the gene chip of the present invention, a general strategy as detailed below can be adopted.

[0058] 1. Slide processing

[0059] First, fully clean the glass slide, use 200mL of 25×NaOH solution and 300mL of 95% ethanol to make 500mL alkaline cleaning solution, thoroughly soak the glass slide in the cleaning solution for 2 hours, then rinse it with water for 5 times and soak the glass slide. In the water. The next step is to soak in poly-L-lysine solution, so that poly-lysine is evenly coated on the glass surface to form a poly-L-lysine coating. The specific method is to make 350mL poly-L-lysine soaking solution, soak the cleaned glass slides directly, keep shaking slowly during soaking, take them out after 1 hour, soak them in water for cleaning, and finally dry them. Generally, the coated glass slides are wrapped with tinfoil and left at room temperature for 1 month for aging to keep ...

Embodiment 2

[0068] Embodiment 2 The general strategy of the gene chip detection sample to be tested according to the present invention

[0069] 1. Labeling of samples to be tested

[0070] According to the different detected objects, select the specific sample to be tested, extract the nucleic acid sample according to the following method, and perform biotin labeling on the nucleic acid of the obtained sample to be tested.

[0071] RNA extraction method: (see Molecular Cloning: Experimental Guide, Chinese Third Edition, 2002, J. Sambrook et al., Huang Peitang et al., pp. 518-522); extract nucleic acid from the sample to be tested.

[0072] Biotin labeling: use the extracted nucleic acid as a template, use RT-PCR method to amplify, use RNA / T7-Oligo (dT) as a primer to synthesize double cDNA, then use cDNA as a template to synthesize cRNA, and use biotin-NTP for labeling .

[0073] 2. Hybridization of the gene chip and the sample to be tested

[0074] Take out the chip and equilibrate th...

Embodiment 3

[0080] Embodiment 3 Transport stress model evaluation chip example

[0081] In this example, the condition of 35°C and 60n / min was used to simulate the transportation stress for 2 hours and continuously for 3 days to study the clinical symptoms, morphological changes of intestinal tissue, cell apoptosis and gene changes in rats after the stress, and through the chip of the present invention for testing and verification.

[0082] 1. Materials and methods

[0083] 1.1 Animal grouping and feeding

[0084] Twenty male SD (sprague-Dawley) rats weighing 250±20 g (purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.). The test rats were adaptively fed for 3 days in an environment with a temperature of 25°C and a humidity of 45% (feeds were given regularly every morning, and water was freely available). They were randomly divided into control group and stress group, with 10 rats in each group. Feed commercial rat food (quantitative 200g per day).

[0085] 1...

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Abstract

The invention relates to a gene chip for evaluating a transport stress model and an application of the gene chip for evaluating the transport stress model. The gene chip comprises one or a plurality of probes used for detecting ribos nucleic acid (RNA), complementary RNA (cRNA) or complementary deoxyribose nucleic acid (cDNA) which is generated by the following genes: Gdnf, Lifr, Ppp3rl, Sphkl, B4galtl, Phldal, Il24, Adm, Gsdmal, Acvrl, Naip2, Aqp2, Ptprc, Bcl2l1, Rela, Ctnnb1, Pdcd6ip, Tgfbr1, Hmox1, Sort1, Tpt1, Aldh1a1, Actc1, Rhot2 and Bax. According to the chip, the typical variations of indexes after livestock and poultry are subjected to transport stress are comprehensively evaluated in a multilayer multi-target point manner through once real-time synchronous high throughput detection of the variation of nerve-immunity-internal secretion indexes, and guiding significance in livestock and poultry organism conditions and production performances during a transport evaluating process is obtained. The chip not only has the characteristics of sample saving and time saving in new drug research, but also saves money and has an important practical meaning.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a gene chip for evaluating transport stress models and its application. Background technique [0002] Transport stress refers to the instinctive adaptation of the animal body under the combined effects of fasting / restricted feeding, environmental changes (mixing, density, temperature, humidity), turbulence, psychological pressure and other stressors during transportation. Sexual and defensive responses are one of the important factors affecting animal production. Under transport stress conditions, animals often show rapid breathing and heartbeat, fear, restlessness, irritability, and a large amount of nutrition and water consumption in the body, which will eventually affect the animal's production performance, immune level and animal product quality. [0003] The mechanism of transport stress is that after the animal is subjected to the action of the stressor, the hypothalamu...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/06
Inventor 许剑琴刘凤华万长荣尹朋麻武仁徐霄龙
Owner CHINA AGRI UNIV
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