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miRNA marker related to pig hair-free character and application of miRNA marker

A marker and trait technology, applied in the field of miRNA markers, can solve the problems of time-consuming and complicated experimental operations, and achieve accurate results and reduce false positives.

Pending Publication Date: 2021-11-30
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Experimental operation is complex and time-consuming

Method used

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  • miRNA marker related to pig hair-free character and application of miRNA marker
  • miRNA marker related to pig hair-free character and application of miRNA marker
  • miRNA marker related to pig hair-free character and application of miRNA marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Screening for the gene miRNA-29a-5p associated with hairless traits

[0041] The experimental population of the present invention is a small amount of large white hairless pigs discovered earlier (discovery address: Anping County, Hengshui City, Hebei Province, discoverer: Ding Xiangdong, associate researcher of the Animal Science and Technology College of China Agricultural University)

[0042] S1. Hairless pig breeding plan: select normal boars and hairless sows to breed, and perform cesarean section when pregnant sows reach 41 days of gestation, take out pig embryos and collect two tissue pieces of size 2cm x 2cm on the back of the embryos, then separate the skin tissue and dry Disinfect. One of them was placed in liquid nitrogen for RNA preservation, and the other was placed in paraformaldehyde for later phenotypic identification.

[0043] S2, phenotype identification: phenotype identification adopts hematoxylin and eosin (HE) staining to carry out hair ...

Embodiment 2

[0054] Example 2 Verifying the relationship between miRNA-29a-5pRNA gene expression and hairless phenotype in pig embryo skin (experimental verification of the relationship between the value of miRNA-29a-5pRNA gene expression and hairless traits)

[0055] S1. Hairless pig breeding plan: select normal boars and hairless sows to breed, and perform cesarean section when pregnant sows reach 41 days of gestation, take out pig embryos and collect two tissue pieces of size 2cm x 2cm on the back of the embryos, then separate the skin tissue and dry Disinfect. Place samples in liquid nitrogen for RNA preservation.

[0056] S2, miRNA-29a-5p primer design:

[0057] The sequence information of the miRNA-29a-5p gene was obtained from the NCBI database (https: / / www.ncbi.nlm.nih.gov / ), and designed using the software miRprimer2. The primers for amplifying the housekeeping gene GAPDH were designed in the same way as above, and were synthesized by Shanghai Sangon Bioengineering Co., Ltd.

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Abstract

The invention relates to a miRNA (micro Ribonucleic Acid) marker related to a pig hair-free character and application of the miRNA marker. The miRNA marker is miRNA-29a-5p, the sequence of the miRNA-29a-5p is shown as SEQ ID NO.1, the precursor sequence of the miRNA-29a-5p is shown as SEQ ID NO.2, and the binding site of the miRNA-29a-5p and a target gene EDAR is shown as SEQ ID NO.3; and when the gene expression quantity of the mature miRNA-29a-5p in the pig embryo skin tissue is lower than 3, the pig is judged as a hairy pig, and when the gene expression quantity of the mature miRNA-29a-5p in the pig embryo skin tissue is higher than 30, the pig is judged as a hair-free pig. The miRNA marker related to the pig hair-free character can be applied to identification and breeding of hair-free pigs, and the hair-free pigs identified and bred according to the technical scheme are applied to the fields of medicine and scientific research.

Description

technical field [0001] The invention relates to the technical field of gene detection and breeding, in particular to a miRNA marker related to pig hairlessness and its application. [0002] technical background [0003] For a long time, pigs have been used as an ideal model animal for medical research to study human medical problems. Hairless pigs have the characteristics of hypoplasia of hair follicles, and hairless pigs can be used as animal models to study diseases in the field of human hair. However, in the process of breeding hairless pigs, how to identify the pigs at the embryonic stage as having hairless traits is rather troublesome. Because the hair shafts in the hair follicles of pig embryos do not protrude from the skin surface, it is impossible to directly determine whether the pig embryos are hairless pigs with the naked eye. The current solution is to first dehydrate the skin tissue of pig embryos to fix the shape, and then conduct histomorphological observation...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/6888C12Q1/6851C12N15/11
CPCC12N15/1138C12Q1/6888C12Q1/6851C12N2310/141C12Q2600/158C12Q2600/124C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 丁向东蒋尧刘华涛邹全李淑娟
Owner CHINA AGRI UNIV