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Method for identifying pure Hareford bulls and hybrid Hareford bulls

A Hereford, purebred technology, applied in the field of animal husbandry, can solve the problem of not identifying the purebred cows of Hereford, and achieve the effects of good social benefits, considerable economic benefits, and strong practicability.

Inactive Publication Date: 2011-01-26
DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the above studies and related literature reports, there is no related research on the promoter region of the TLR2 gene, and there is no use of specific alleles in the promoter region of the LTR2 gene to identify Hereford purebred cattle, Hereford crossbred cattle and other breeds cow report

Method used

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  • Method for identifying pure Hareford bulls and hybrid Hareford bulls
  • Method for identifying pure Hareford bulls and hybrid Hareford bulls
  • Method for identifying pure Hareford bulls and hybrid Hareford bulls

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Experimental program
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Embodiment 1

[0021] Genomic DNA was extracted from the first target bovine blood by phenol-chloroform extraction, and specific primers were designed to amplify the TLR2 gene promoter region fragment. The primer sequences used were:

[0022] The upstream primer sequence is: SEQ ID NO.4:

[0023] TLR2-promoter-F: 5'-ggattctccaggcaagaaca-3';

[0024] The downstream primer sequence is: SEQ ID NO.5:

[0025] TLR2-promoter-R: 5'-agtcctgtggtggtcccttt-3'.

[0026] The PCR reaction system used was as follows: the total system was 20 microliters (μl), including 10-100 ng of genomic DNA, 1.0 μl of upstream primers (10 pmol / μl), 1.0 μl of downstream primers (10 pmol / μl), 2.0 μl of 10 ×PCR Buffer (containing magnesium ions Mg 2+ ), 2 μl of dNTPs (0.25 mM), 2.0 U of Taq DNA polymerase and sterilized water.

[0027] The PCR reaction program used was as follows: pre-denaturation at 94°C for 4 min; 35 cycles of denaturation at 94°C for 30 s, annealing at 60°C for 30 s, and extension at 72°C for 45 s; e...

Embodiment 2

[0030] Genomic DNA was extracted from the second target bovine blood by phenol-chloroform extraction, and specific primers were designed to amplify the TLR2 gene promoter region fragment. The primer sequences used were:

[0031] The upstream primer sequence is: SEQ ID NO.4:

[0032] TLR2-promoter-F: 5'-ggattctccaggcaagaaca-3';

[0033] The downstream primer sequence is: SEQ ID NO.5:

[0034] TLR2-promoter-R: 5'-agtcctgtggtggtcccttt-3'.

[0035] The PCR reaction system used was as follows: the total system was 20 microliters (μl), including 10-100 ng of genomic DNA, 1.0 μl of upstream primers (10 pmol / μl), 1.0 μl of downstream primers (10 pmol / μl), 2.0 μl of 10 ×PCR Buffer (containing magnesium ions Mg 2+ ), 2 μl of dNTPs (0.25 mM), 2.0 U of Taq DNA polymerase and sterilized water.

[0036] The PCR reaction program used was as follows: pre-denaturation at 94°C for 4 min; 35 cycles of denaturation at 94°C for 30 s, annealing at 60°C for 30 s, and extension at 72°C for 45 s; ...

Embodiment 3

[0039] Genomic DNA was extracted from the blood of the third target bovine by phenol-chloroform extraction, and specific primers were designed to amplify the TLR2 gene promoter region fragment. The primer sequences used were:

[0040] The upstream primer sequence is: SEQ ID NO.4:

[0041] TLR2-promoter-F: 5'-ggattctccaggcaagaaca-3';

[0042] The downstream primer sequence is: SEQ ID NO.5:

[0043] TLR2-promoter-R: 5'-agtcctgtggtggtcccttt-3'.

[0044] The PCR reaction system used was as follows: the total system was 20 microliters (μl), including 10-100 ng of genomic DNA, 1.0 μl of upstream primers (10 pmol / μl), 1.0 μl of downstream primers (10 pmol / μl), 2.0 μl of 10 ×PCR Buffer (containing magnesium ions Mg 2+ ), 2 μl of dNTPs (0.25 mM), 2.0 U of Taq DNA polymerase and sterilized water.

[0045] The PCR reaction program used was as follows: pre-denaturation at 94°C for 4 min; 35 cycles of denaturation at 94°C for 30 s, annealing at 60°C for 30 s, and extension at 72°C for ...

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Abstract

The invention provides a method for identifying pure Hareford bulls and hybrid Hareford bulls, which is to determine if a Hareford bull is pure or hybrid or of other variety by detecting the loci from 300bp to 307bp in a promoter area 627bp of a TLR2 gene of the bull. The method has the characteristics of simplicity, high speed, accuracy, reliability, low cost and the like and has high practicality. The method of the invention has a promising application prospect in Hareford bull introduction, Hareford import and export trade, local crossbreeding improvement, beef cattle novel variety breeding, beef product molecular marking and tracking and the like.

Description

technical field [0001] The invention relates to the technical field of animal husbandry, in particular to a method for identifying Hereford special purebred cattle, Hereford crossbred cattle and other breeds of cattle by using a TLR2 (Toll-like receptor2) gene-specific promoter. Background technique [0002] According to the survey data of "Status of China's Livestock and Poultry Genetic Resources" published in 2003, there are 69 breeds of cattle in my country, including 52 local breeds, 5 cultivated breeds, and 12 introduced breeds. It is the country with the most breeds of cattle in the world. Chinese native yellow cattle have been mainly used for draft for a long time, and they have the advantages of good meat quality, resistance to rough feeding, and strong stress resistance (Qiu Huai et al., 1986). However, the vast majority of cattle breeds in my country do not have the body structure characteristics of good beef cattle breeds, and have disadvantages such as low meat p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 黄金明刘利鞠志花王长法王泽英李建斌李秋玲李荣岭侯明海仲跻峰
Owner DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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