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Primer, method and kit for quick screening of curly-hair genes of cattle

A kit and gene technology, applied in the field of animal husbandry, can solve the problems of declining milk production and reproductive performance, affecting economic benefits, and stunted growth and development of cattle, so as to reduce the probability of parasitic diseases and improve the economic benefits and costs of breeding. low cost effect

Inactive Publication Date: 2016-11-16
DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the actual production of the cattle industry, curly-haired cattle are more likely to be parasitic with ticks and other parasites, resulting in stunted growth, reduced milk production and reproductive performance of cattle, affecting their economic benefits

Method used

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  • Primer, method and kit for quick screening of curly-hair genes of cattle
  • Primer, method and kit for quick screening of curly-hair genes of cattle
  • Primer, method and kit for quick screening of curly-hair genes of cattle

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Effect test

Embodiment 1

[0032] 1.1 Genomic DNA was extracted from bovine blood by phenol-chloroform extraction, and the DNA sample was diluted to 50 ng / μL for later use. A total of 75 cattle were used in the experiment, including 18 Altay white-headed cattle, 20 Nanyang cattle, 17 Xinjiang brown cattle, and 20 Luxi yellow cattle.

[0033]1.2 According to the gene sequence of KRT27 with Genbank accession number AC_000176.1 (41631641..41637275), use Primer Premier5.0 software to design specific primers for amplifying the bovine KRT27 gene fragment. Exon 1 of the KRT gene is 443bp in total. When designing primers, attention should be paid to the length of the amplified fragment greater than 443bp, the length of the primer is 15-30bp, the GC content is 40%-60%, and the Tm is between 55-80°C, while avoiding the product secondary structure area. In summary, a pair of primers specially designed for KRT gene amplification, the length of the amplified product is 469bp, the length of the primer is 20bp, the G...

Embodiment 2

[0057] 2.1 Genomic DNA was extracted from the blood of 23 Simmental cattle by phenol-chloroform extraction. Utilize the PrimerPremier5.0 software to design specific primers for amplifying the bovine KRT27 gene fragment, and the primer sequences used are:

[0058] The upstream primer sequence is: SEQ ID NO.1:

[0059] 5'-GAGCGTGCGCTTTTCTTCTG-3'

[0060] The downstream primer sequence is: SEQ ID NO.2:

[0061] 5'-TAAACCTCACGGCACTGTGT-3'

[0062] 2.2 According to the above designed primers for detecting bovine curly hair gene carriers, the following steps are used to amplify the bovine KRT27 gene fragment, and the amplified fragment is detected by agarose gel electrophoresis with a mass concentration of 1%.

[0063] (1) Use 25 μl of PCR reaction system: DNA (100ng / μl) 1 μl, upstream and downstream primers (10 μmol / L) 0.5 μl each, 2×Taq PCR MasterMix 12.5 μl, double distilled water (ddH 2 O) Make up to 25 μl.

[0064] (2) PCR reaction conditions: denaturation at 94°C for 4 mi...

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Abstract

The invention discloses a primer, a method and a kit for quick screening of curly-hair genes of cattle, and belongs to the technical field of stock farming. The method includes using a genome DNA of cattle to be detected as a template, designing upstream and downstream primers on KRT27 genes for PCR amplification, subjecting PCR products to enzyme digestion with Takara restriction enzyme AvaII after reaction, conducting polyacrylamide gel electrophoresis after enzyme detection, and detecting that cattle with 469bp as DNA segments is uncurly-hair gene cattle; and detecting that cattle with 195bp, 274bp and 469bp as DNA segments are curly-hair gene heterozygote cattle. With the primer, curly hair of calves can be identified in the early stage, parasite risk is reduced for selected cattle, and high economic value is achieved. Meanwhile, hair curling as visual phenotype information can be used as a genetic marker and provide guidance for genetic breeding of cattle.

Description

technical field [0001] The invention belongs to the technical field of animal husbandry, and in particular relates to a primer, a method and a kit for rapid screening of bovine curly hair genes. Background technique [0002] Skin tissue is a natural barrier for animals to protect themselves from foreign objects. The formation of skin promotes the development of hair follicles, and hair is a keratinized product produced by hair follicles in skin tissue. The formation of hair fibers is regulated by the interaction of hair follicle epidermal cells and mesenchymal cells, and only occurs in the early stage of the hair growth cycle, stops in the catagen phase, and finally stops in the telogen phase. The phenotypic characteristics of hair, including the length, density, shape and color of hair, will show different characteristics with different parts of the animal body. Different species, or different species of the same species, have great differences in coat phenotype, which is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 姜强黄金明王秀革张燕鞠志花王长法孙艳仲跻峰
Owner DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI