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Method for assisting judgment of Wuzhishan minipig inbred line

A technology for assisting identification and miniature pigs, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc.

Inactive Publication Date: 2009-12-23
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to various reasons, it has not been realized. The key is that West Germany does not easily send resources to China through cooperative research.

Method used

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  • Method for assisting judgment of Wuzhishan minipig inbred line
  • Method for assisting judgment of Wuzhishan minipig inbred line
  • Method for assisting judgment of Wuzhishan minipig inbred line

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, the discovery of Pnas-4 gene C1400T polymorphism

[0022] The pigs used in the experiment were as follows: 13 Landrace pigs, 19 Large White pigs, 45 Tongcheng pigs, 42 Guizhou miniature pigs, 49 Guangxi Bama miniature pigs and 42 Wuzhishan miniature pig inbred lines.

[0023] 1. PCR amplification

[0024] A pair of primers were designed according to the second intron and the 3'UTR sequence of the Pnas-4 gene (GenBank Accession Number DQ406743) (sequence 1 of the sequence listing) as follows:

[0025] Upstream primer (sequence 2 of the sequence listing): 5'-CTAGAACCACTCAAACCAAGCAGC-3';

[0026] Downstream primer (sequence 3 of the sequence listing): 5'-ATCAGGCAGGTAAAAGGATAACGG-3'.

[0027] Utilizing the above primer pairs, PCR amplification was carried out using pig genomic DNA as a template.

[0028] PCR reaction system: 2.0 μL 10x reaction buffer, 1.6 μL MgCl 2 (2.5mmol / L), 1 μL upstream primer (10 μmol / L), 1 μL downstream primer (10 μmol / L), 0.4 μL d...

Embodiment 2

[0047] Embodiment 2, the application of Pnas-4 gene C1400T polymorphism

[0048] The 75 ear skin tissue samples were all collected from the Wuzhishan miniature pig inbred line of the Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences, of which F 13 Generation 15 heads, F 14 Generation 13 heads, F 15 Generation 11 head, F 16 Generation 14 heads, F 17 Generation 14 heads, F 18 Generation 8, all samples were treated with 75% alcohol and stored in -20°C refrigerator for DNA extraction.

[0049] 1. PCR amplification

[0050] Primers are as follows:

[0051] Upstream primer: 5′-CTAGAACCACTCAAACCAAGCAGC-3′;

[0052] Downstream primer: 5'-ATCAGGCAGGTAAAAGGATAACGG-3'.

[0053] Utilizing the above primer pairs, PCR amplification was carried out using pig genomic DNA as a template.

[0054] PCR reaction system: 2.0 μL 10x reaction buffer, 1.6 μL MgCl 2 (2.5mmol / L), 1 μL upstream primer (10 μmol / L), 1 μL downstream primer (10 ...

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Abstract

The invention discloses a method for assisting the judgment of a Wuzhishan minipig inbred line. The method for assisting the judgment of the Wuzhishan minipig inbred line and / or a non-Wuzhishan-mountain minipig inbred line, which is provided by the invention, detects that the 1400th position of deoxyribonucleotide from the 5' end of GenBankAccession Number DQ406743 of a pig to be detected is C or T, and determines that the genotype of the pig to be detected is CC, CT or TT; the pig to be detected with the CC genotype is an alternate Wuzhishan minipig inbred line; and a pig with the CT genotype and a pig to be detected with the TT genotype are non-Wuzhishan-minipig inbred lines. The method can be applied to the seed breeding of the Wuzhishan minipig inbred line. The method can firstly carry out preliminary screening on all the pigs in a swinery to be detected, eliminates the non-Wuzhishan-minipig inbred line, finds out the alternate Wuzhishan minipig inbred line and combines with other methods to carry out further validation. The invention can also be used for detecting whether the Wuzhishan minipig inbred line purchased in the market is counterfeit or not.

Description

technical field [0001] The invention relates to a method for assisting identification of inbred lines of Wuzhishan miniature pigs. Background technique [0002] In the early days of my country's reform and opening up, when researchers were studying in West Germany from 1983 to 1984, they had the honor to visit the Gottengen Minieture Pig Breeding Center in West Germany and learned that miniature pigs have been widely used in animal models of human diseases in Western countries. At the same time, it is an excellent research object for pig embryo engineering. Göttingen minipigs have been sold to many countries and regions, so the idea of ​​bringing them back to China for development and application came into being. Professor Schmidt, one of the miniature pig breeding experts in Göttingen, West Germany, is interested in the Chinese Meishan pig. He suggested that the West German Ministry of Agriculture list it as a special support for bilateral cooperative research, and the resea...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11A01K67/027
Inventor 冯书堂杨述林李奎牟玉莲
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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