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Pouch pig SLA-2 genes and application thereof

A technology of SLA-2 and purse pig, which is applied in application, genetic engineering, plant gene improvement, etc., can solve the problems of commercial breeding of purse pig, lack of scientific and effective guidance for breed identification and artificial breeding, etc.

Inactive Publication Date: 2012-06-06
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is a lack of scientific and effective guidance for its variety identification and artificial breeding, which has brought huge problems to the commercial breeding of purse pigs

Method used

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  • Pouch pig SLA-2 genes and application thereof
  • Pouch pig SLA-2 genes and application thereof
  • Pouch pig SLA-2 genes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Cloning of SLA-2 alleles of piglets

[0029] 1. Primer design

[0030] Utilize RT-PCR to amplify the above-mentioned pouch pig SLA-2 allele, the primer sequence used for amplification is:

[0031] Upstream primer S1: 5'-AGATGCGGGTCAGGGGCCCTCAAG-3' (SEQ ID NO: 5);

[0032] Downstream primer S2: 5'-CAGTCCCCACAAGGC AGCTGTCTC-3' (SEQ ID NO: 6);

[0033] 2. Extraction of total RNA from porcini

[0034] The spleen of the purse pig was collected, and the total RNA of the nucleic acid was extracted using the TRIZOL kit (BS409, Canada Bio Basic Inc.), and the method was carried out according to the kit instructions.

[0035] 3. RT-PCR

[0036] Using Oligo dT as a reverse transcription primer, reverse-transcribe the nucleic acid into cDNA according to the instructions of the reverse transcription kit (M-MLV, RNase H-, Treasure Bioengineering (Dalian) Co., Ltd.); then use PCR to amplify primer S1 / S2 amplifies SLA-2.

[0037] The amplification RT-PCR program is: ...

Embodiment 2

[0048] Embodiment 2: Sequence analysis and identification of varieties

[0049] The gene sequence obtained in Example 1 was analyzed through Gentyx9.0 (Software Development co., Ltd), and its amino acid sequence was deduced; the amino acid sequences of 44 SLA-2 alleles except the purse pig were mobilized on DDBJ / EMBL / GenBank , amino acid sequence comparison using DNAMAN Version5.2.2 (Lynnon Biosoft).

[0050]The analysis results are attached Figure 1~4 Shown: The 47th, 48th, 67th, 68th, 74th, 97th, 119th, 138th, and 240th positions of the SLA-2 gene in the coding region of the purse pig are characteristic amino acids, and the specific information is as follows :

[0051] Location

amino acid

English name of amino acid

amino acid abbreviation

47

Phenylalanine

Phenylalanine

F or Phe

48

Isoleucine

Isoleucine

I or Ile

67

Alanine

Alanine

A or Ala

68

Lysine

Lysine

K or Lys

...

Embodiment 3

[0054] Example 3: Drawing of Molecular Evolution Atlas and Its Application in Optimizing Genetic Breeding

[0055] Molecular evolution maps were drawn using the neighbor-joining method of DNAMAN and Mega2 software (Mega software Co., Ltd). The molecular evolution map is attached Figure 5 . According to the molecular evolution map, all SLA-2 alleles were clustered into three categories, namely, I, II and III, and the SLA-2 alleles of the purse pig belonged to the B III region. From this, it can be clearly judged that the purse pig strain is close to the American Hanford AF464039, the American Large White EF125047, and the cell line 10sk21 EU432083; while it is genetically related to the Chinese hybrid strain AF205147, the American NIH miniature pig EU867814, and the American CMS miniature pig EU431219. The relationship is far away. According to the principle of genetic breeding, under the condition of the same dominant traits, we should try our best to select the strains wit...

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Abstract

The invention relates to pouch pig SLA-2 genes and application thereof. The application comprises amplification, sequence determination and analysis of four SLA-2 allelic genes cDNA of pouch pigs of a Chinese characteristic variety. The base sequences of the four SLA-2 allelic genes are expressed as SEQ ID No: 1, SEQ ID No: 2, SEQ ID No: 3 and SEQ ID No: 4. The gene sequences and genetic molecular characteristics and molecular evolution maps obtained based on the gene sequence can be used for construction of a pouch pig gene bank, variety identification of the pouch pigs and guidance on genetic breeding of the pouch pigs.

Description

technical field [0001] The invention relates to the amplification, sequence determination and analysis of the cDNA of the purse pig SLA-2 allele (SLA-2-HB) cDNA of a Chinese characteristic variety, and the use of the purse pig variety identification and optimized genetic breeding. Background technique [0002] The purse pig is a small group of Northeast folk pigs, which are distributed in the closed mountainous area of ​​western Liaoning. It is named for its shape resembling a "purse". This breed of pigs has been self-breeding and self-breeding for a long time, and has evolved naturally for more than 300 years. This breed of pigs is wild and has good disease resistance. In addition, this breed of pork is delicious and mellow in taste. It is commonly known as "Northern Fragrant Pig". known as. In 1987, it was listed as a national first-class protected pig breed by the Ministry of Agriculture. In 2002, it was included in the "National Livestock and Poultry Genetic Resources P...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/10C12Q1/68
Inventor 高凤山许崇波
Owner DALIAN UNIV
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