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Pig isocitric acid dehydrogenase gene IDH3-gamma as genetic marker of production trait in pigs

A kind of isocitrate dehydrogenase, gene technology, applied in the field of molecular marker-assisted selection of pigs

Inactive Publication Date: 2008-04-09
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007]However, the cDNA full-length cloning, genome structure analysis and polymorphism research of porcine IDH3γ gene are still blank at home and abroad

Method used

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  • Pig isocitric acid dehydrogenase gene IDH3-gamma as genetic marker of production trait in pigs
  • Pig isocitric acid dehydrogenase gene IDH3-gamma as genetic marker of production trait in pigs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of cDNA sequence of isocitrate dehydrogenase IDH3γ gene

[0026] The preparation of the full-length cDNA sequence of porcine isocitrate dehydrogenase IDH3γ gene needs to first synthesize the forward cDNA duplex (use the kit from Clontech Company in the United States, and operate according to the operation manual of the kit). During the transcription process, the reverse transcriptase has more than 3 bases G connected to the 5' end of the cDNA single strand, and at the same time, the linker primer is connected to the two ends of the cDNA sequence, and then the LD-PCR is carried out with the head primer ( Long Distance PCR) to obtain double-stranded full-length cDNA. The three primers used to prepare the forward cDNA duplex were: forward TII oligonucleotide primer: 5'-AAGCAGTGGTATCAACGCAGAGTACGCGGG-3' (12 μM), CDS primer: 5'-AAGCAGTGGTATCAACGCAGAGTACT(30)VN-3'( 12 μM), PCR primer: 5′-AAGCAGTGGTATCAACGCAGAGT-3’ (12 μM). The brief steps for synthesi...

Embodiment 2

[0033] Example 2: The acquisition of gene fragments and the establishment of polymorphism detection methods

[0034] Choose 2 large white pigs (foreign ancestry) and 2 Meishan pigs (Chinese local pig ancestry) according to the genomic structure of human IDH3γ gene and the cDNA sequence of pig IDH3γ gene obtained by Example 1 as test materials, design the following primers, the primers as follows:

[0035] Forward primer F1: 5'-GTTCTAGGTGCCCACGAG-3';

[0036] Reverse primer R1: 5'-TTGACATGCAGCATGAGC-3'.

[0037] Use these primers to carry out PCR amplification in the genomic DNA of 2 large white pigs and 2 Meishan pigs. The PCR reaction system is 25 μl, wherein the template DNA is 50ng, the concentration of dNTPs is 200 μmol / L, and the concentration of each primer is 0.3 μmol / L. 1U of Taq DNA polymerase (Biostar International, Canada), added deionized water to a total volume of 25 μl; PCR reaction program: pre-denaturation at 94°C for 4 minutes; then denaturation at 94°C for ...

Embodiment 3

[0043] Embodiment 3: Detection of polymorphic distribution of genetic markers of the present invention in different pig herds

[0044] The microsatellite polymorphisms in the second intron of porcine IDH3γ gene were detected in 2 commercial pig herds (Large White and Landrace) and 6 Chinese local pig breeds (Meishan, Huainan, Bamei, Tongcheng, Hezuo and Erhualian). The test results are shown in Table 1. The A allele was dominant in several pig herds tested, and the B allele was not found in Changbai, Meishan, Hezuo and Erhualian pigs.

[0045] Table 1 The genotype and genotype frequency of the porcine IDH3γ gene microsatellite polymorphism prepared by the present invention in different pig groups

[0046]

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Abstract

The invention pertains to the porcine marker-assisted selection technical field. A genetic mark which is suitable for detecting porcine productive trait is cloned by the invention. The genetic mark is porcine isocitric acidcitric dehydrogenase gene IDH3Gamma, and cDNA sequence of the gene is stated in sequence table SEQID NO: 1. the sequence length is 1346bp, containing an open reading frame of 1179bp, an untranslated region of 36bp5' and a 3' untranslated region of 131p. The nucleotide sequences of IDH3Gamma gene such as SEQID NO: 3, SEQID NO: 4, SEQID NO: 5 and SEQID NO: 6 contain a second intron sequence of IDH3Gamma, and provide a new genetic mark the porcine marker-assisted selection. The invention also discloses a cDNA sequence of porcine IDH3Gamma gene and DNA sequence of the second intron for application of the pig microsatellite typing test.

Description

technical field [0001] The invention belongs to the technical field of molecular marker-assisted selection of pigs, and in particular relates to the cloning of porcine isocitrate dehydrogenase gene IDH3γ gene and its application as a genetic marker in marker-assisted selection of pig production traits. Background technique [0002] The genetic improvement of livestock production traits has been greatly facilitated by selective breeding of animals based on breeding values ​​estimated from phenotypic values ​​of production traits. Since the 1990s, with the advancement of molecular biology technology and its application in the field of pigs, molecular marker-assisted selection and infiltration molecular breeding techniques with molecular markers as the core have gradually appeared. Accelerated the pig breeding process. Genes or markers that can be applied to molecular marker-assisted selection must have a large genetic contribution rate to the target trait, that is, the main e...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/53
Inventor 熊远著任竹青雷明刚左波
Owner HUAZHONG AGRI UNIV
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