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Screening expression profile of growth specific genes in swine and functional cDNA chip prepared by using the same

A functional and specific technology, applied in the field of screening growth-specific genes, can solve the problems of cost-intensive work, lack of genes, difficulties, etc.

Inactive Publication Date: 2005-06-01
庆尚南道 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the role of non-related genes, it will encounter difficulties when converting the composite DNA chip to reagent use, and it will take a lot of work to finally interpret the biological role of the genes
At the same time, since it is based on database information, the difficulty lies in the lack of information on new genes or possibly related genes to a certain extent

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Screening for expression patterns of growth-related specific genes in pigs

[0020] To screen the expression patterns of specific genes associated with pig growth, probe DNA was prepared from total RNA isolated from muscle and adipose tissue of Kagoshima black pigs (Kagoshima Berkshire), and the total RNA in these tissues was analyzed. Fluorescent labeling to prepare a target DNA. These DNAs are hybridized and scanned. The obtained image data were analyzed to screen pig growth-related specific genes, which were then cloned to determine the nucleotide sequence.

preparation Embodiment 1

[0021] Preparation Example 1: Preparation and Arrangement of Probe DNA

[0022] First, probe DNA, which is cDNA amplified by PCR, is prepared and attached to a glass slide. Total RNA was extracted from the longissimus dorsi muscle and adipose tissue of Kagoshima black pigs (body weight 30kg and 90kg) with RNA extraction kit (Qiagen, Germany) according to its instructions, and oligo(dT) column Extract mRNA. The extracted mRNA samples were subjected to RT-PCR to synthesize cDNA using SP6, T3 forward primer, T7 reverse primer (Amersham Pharmacia Biotech, England). The total volume of each PCR reaction was 100 μl. 100 pM of forward and reverse primers were transferred separately to 96-well PCR plates (Genetics, England). Each well contains 2.5mM dNTP, 10×PCR buffer, 25mM MgCl 2 , 0.2 μg of DNA template, 2.5 units of Taq polymerase. PCR was performed with GeneAmp PCR System 5700 (AB Applied BioSystem, Canada), under the following conditions: 94°C, 30 seconds; 58°C, 45 seconds...

preparation Embodiment 2

[0025] Preparation Example 2: Preparation and Hybridization of Target DNA

[0026] To prepare target DNA for screening growth-related genes in pig muscle and adipose tissue, muscle tissue was obtained from the longissimus dorsi of Kagoshima black pigs weighing 30 kg and 90 kg, and adipose tissue was obtained from the longissimus dorsi muscle of Kagoshima black pigs weighing 30 kg Kagoshima black pigs. Muscle and adipose tissue were cut into 5-8mm lengths and stored in liquid nitrogen at -70°C.

[0027] According to Trizol TM According to the kit (Life Technologies, Inc.), total RNA was isolated from 0.2 to 1.0 g of tissues of the experimental group and control group to prepare target DNA. Trizol TM Add to tissue, Trizol TM The dosage is to add 1mL Trizol per 50~100mg tissue TM , use a glass-Teflon or Polytron homogenizer to disrupt the tissue, then centrifuge at 12,000 g for 10 minutes at 4°C, and take a 1 mL supernatant sample for testing. Add 200 μl of chloroform to ...

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Abstract

The present invention relates to screening the expression patterns of growth-specific genes of pigs and functional cDNA chips using these genes, and provides the nucleotide sequences of new growth factors related to growth in pig muscle and adipose tissue. At the same time, the present invention provides a functional cDNA chip for screening and functional analysis of growth-specific genes according to pig breeds and tissues. integrated in it. Therefore, the functional cDNA chip for screening and functional analysis of growth-specific genes according to pig breeds and tissues can be used to improve pig quality and breed new breeds.

Description

technical field [0001] The present invention relates to the screening of expression patterns of growth-specific genes in pigs and functional cDNA chips using these genes. More specifically, the present invention relates to the screening of expression patterns of growth-specific genes that are specifically expressed in muscle and fat tissue of growing pigs, and also relates to a method for screening pigs according to their breed and tissue. A functional cDNA chip for screening and functional analysis of growth-specific genes, the cDNA chip is prepared by integrating only the specific genes obtained by the screening into it. Background technique [0002] The development of molecular biology has had a huge impact on the field of livestock genetics and breeding, and has made great achievements in the study of genetic linkage map (genetic linkage map) and quantitative trait loci (quantitative trait loci (QTL) map) of pigs Great progress. In particular, the mapping of QTLs assoc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6837C12Q1/6888C12Q2600/158C12Q1/68
Inventor 金哲旭吕政秀李正圭宋瑛敏曹光根郑起和金一锡陈祥根朴修贤郑智媛李旼晶权银贞赵恩锡赵碻来慎诜敏南喜善洪娟喜洪星光姜阳守河永主卢正晚郭石俊崔仁灏金炳宇
Owner 庆尚南道
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