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ABCF1 gene used as porcine birth weight character related molecular marker

A technology of molecular markers and traits, applied in the field of molecular marker-assisted selection of pigs, can solve the problem of lack of transmembrane domains

Inactive Publication Date: 2013-09-11
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the protein lacks the transmembrane domain of most ABC transporters

Method used

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  • ABCF1 gene used as porcine birth weight character related molecular marker
  • ABCF1 gene used as porcine birth weight character related molecular marker
  • ABCF1 gene used as porcine birth weight character related molecular marker

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Embodiment 1: Porcine ABCF1 gene partial DNA sequence amplification

[0019] (1) Extraction of total DNA from large white pig (Landrace pig) muscle tissue by phenol extraction

[0020] 1) Grind the muscle tissue of large white pigs (or Landrace pigs, from the boutique pig farm of Huazhong Agricultural University in Wuhan, Hubei Province, which is a lean pig breed of foreign blood that is routinely promoted) in liquid nitrogen, and add an equal volume of 1× SET solution (1 ml), proteinase K (10 mg / mL) to a final concentration of 200 μg / mL, sodium dodecyl sulfate (ie SDS, 10%) to a final concentration of 0.5%, and shake well. Incubate overnight in a shaker at 55°C.

[0021] 2) Add an equal volume of Tris-saturated phenol to the digested tissue sample, slowly invert the centrifuge tube for 15 minutes, centrifuge in a low-temperature refrigerated centrifuge at 4°C and 11,000 rpm for 10 minutes, and carefully transfer the supernatant to another centrifuge tube. on the corr...

Embodiment 2

[0037] Embodiment 2: The establishment of PCR-RFLP diagnostic method

[0038] (1) PCR-RFLP primer sequence (this primer is also the primer for amplifying the 16th intron of ABCF1 gene), the sequence of this primer pair is as follows:

[0039] ABCF1 SNP: forward primer 5'-TGGGGTGAGTTAAGGCTG-3', (corresponding to figure 2 1-18 of the sequence shown),

[0040] Reverse primer 5'-GGGCTTGGTTGGTTTGC-3'. (corresponding to figure 2 581-597 of the sequence shown);

[0041] The length of the primer amplified fragment is 597bp (see figure 2 and Sequence Listing SEQ ID NO: 1).

[0042] (2) PCR amplification conditions

[0043] The total volume of the PCR reaction is 10 μl, including 1 μl of the porcine genomic DNA template prepared above, 7.1 μl of double distilled water, 1 μl of 10× buffer, 0.3 μl of each 10 mM forward primer and reverse primer, 0.2 μl of dNTP, and 0.1 μl of Taq enzyme (5 U / μl). The PCR reaction conditions were: 94°C pre-denaturation for 5 minutes, followed by 35...

Embodiment 3

[0047] Example 3: Application of the molecular markers of the present invention in correlation analysis of pig reproductive traits 1

[0048] The pigs in this example come from the Landrace pigs (a breed that is routinely popularized and applied) in the Shuitai original breed pig farm of Guangdong Huanong Wen's Animal Husbandry Co., Ltd., and the weight of the piglets was measured on day 0 after birth.

[0049] According to the population structure of the collected samples, the applicant used a mixed model to statistically analyze the genotype effect of the A / G polymorphism site in the 16th intron region of the ABCF1 gene and its relationship with the traits of birth weight. The fixed effect includes the sex effect , parity effect, line effect, genotype effect of candidate genes, random effects including sire effect, sire effect within sire; MIXED MODELS program in SAS (VersioB 8.0) software was used for least square mean estimation (Breslow, N.E.; Clayton, D.G. Approximate In...

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Abstract

The invention belongs to the technical field of preparation of livestock molecular markers, and particularly relates to a birth weight character related molecular marker used for porcine marker auxiliary selection and application thereof. The molecular marker is prepared by cloning an ABCF1 gene, and the DNA (deoxyribonucleic acid) sequence is disclosed as SEQ ID NO:1 in the sequence table and as Figure 2. The 256bp position of the SEQ ID NO:1 in the sequence table has an A256-G256 base mutation, resulting in MspI-RFLP polymorphism. The invention also discloses primers for amplifying partial sequence of the 16th introne of the ABCF1 gene and a polymorphism detection method by using the same. The invention provides a new molecular marker for porcine marker auxiliary selection.

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 and application of molecular markers related to birth weight traits used as marker-assisted selection of pigs. The molecular marker of the present invention is related to ABCF1 gene. Background technique [0002] Pig is an important economic animal. Pork has been favored by consumers for a long time because of its tender and delicious meat. It is one of the main sources of animal protein for our people. In recent years, people's consumption of pork has been increasing day by day. How to improve production performance and reduce production costs has become one of the priorities of pig breeders. [0003] The rapid development of molecular biology technology has provided an important opportunity for this. With this technology, scientists have discovered a large number of molecular markers that are significantly associated ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 余梅李小平赵书红梁跃朱猛进李长春李新云曹建华
Owner HUAZHONG AGRI UNIV
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