Pouch pig SLA-1 gene and application thereof

A technology of SLA-1 and purse pigs, which is applied in application, genetic engineering, plant gene improvement, etc., can solve the problems of commercial breeding of purse pigs, lack of scientific and effective guidance for breed identification and artificial breeding, etc.

Inactive Publication Date: 2011-02-23
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 an

Method used

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  • Pouch pig SLA-1 gene and application thereof
  • Pouch pig SLA-1 gene and application thereof
  • Pouch pig SLA-1 gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Cloning of the SLA-1 allele of the purse pig

[0029] 1. Primer design

[0030] Utilize RT-PCR to amplify the above-mentioned pouch pig SLA-1 allele, the primer sequence used for amplification is:

[0031] Upstream primer pM-1F: 5'-ATGGGGCCTGGAGCCCTCTTC-3' (SEQ ID NO: 5);

[0032] Downstream primer pM-1R: 5'-TTAGGATGCTCATAGGAACACA-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, Bio Basic Inc., Canada), 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 the primer pM -1F / pM-1R amplifies SLA-1.

[0037] The amplification ...

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 58 other SLA-1 alleles except the purse pig were mobilized on DDBJ / EMBL / GenBank , amino acid sequence alignment using DNAMAN Version 5.2.2 (Lynnon Biosoft).

[0050] The analysis results are attached Figure 1~4 Shown: The 88th, 89th, 90th, 92nd, 97th, 135th, 145th, 176th and 177th positions of the SLA-1 gene in the coding region of the purse pig are characteristic amino acids, the specific information is as follows :

[0051] Location

amino acid

English name of amino acid

amino acid abbreviation

88

Glutamine

Glutamine

Q or Gln

89

arginine

Arginine

R or Arg

90

aspartic acid

Aspartic acid

D or Asp

92

serine

Serine

S or Ser

...

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 . Through the molecular phylogenetic tree, it can be clearly judged that the genetic distance between the purse pig strain and American Sinclair pig AF464045, Meishan pig AY135600, etc. is relatively close, while the genetic distance with Korean strains DQ883208, DQ992489, etc. is relatively far. According to the principle of genetic breeding, under the condition of the same dominant traits, we should try our best to select the strains with distant genetic relationship to reflect the hybrid advantage; and in order to preserve the species, we should try our best to select the strains with closer genetic relationship.

[0056] Based on the above information and breeding principles, Jianchang C...

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Abstract

The invention discloses a pouch pig SLA-1 gene and application thereof. The invention comprises cDNA amplification, sequencing and analysis of a pouch pig in characteristic species in China which comprises four SLA-1 alleles, wherein the four SLA-1 alleles have the following base sequences shown as SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4. The gene sequences and gene molecular characteristics and a molecular evolution map obtained on the basis can be used for constructing a gene library of the pouch pig, identifying species of the pouch pig and guiding inheritance breeding of the pouch pig.

Description

technical field [0001] The invention relates to the amplification, sequence determination and analysis of the cDNA of SLA-1 allele (SLA-1-HB) cDNA of a special variety of purse pig in my country, as well as the identification and optimized genetic breeding of purse pig species by using it. 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 Re...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/12C12N15/10
Inventor 高凤山白婧
Owner DALIAN UNIV
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