Molecular Markers and Their Application in Breeding of Holstein Cattle for Resistance to Heat Stress
A technology of Holstein cattle and heat stress resistance, applied in the field of molecular genetics, can solve the problems of the application of heat shock protein 70, etc., achieve the effects of convenient and fast detection, early auxiliary selection, and low detection cost
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Embodiment 1
[0029] 1 DNA extraction
[0030] Take the blood sample, tissue sample or milk sample (choose one of the three samples) of the dairy cow to be tested, according to Jin Dongyan, etc. [1] Wada rain [2] method to extract DNA from the sample.
[0031] 2 Apply the primers SEQ ID No.1 and SEQ ID No.2 screened by the present invention to carry out PCR-SSCP analysis on the extracted DNA and standard DNA, and carry out non-denaturing polyacrylamide electrophoresis to the amplified product, and analyze the electrophoretic pattern To carry out gene polymorphism analysis, the specific operation process is as follows:
[0032] Use the above primers to carry out PCR amplification on the PE 9600 amplification instrument: the PCR reaction volume is 12.5 μl, of which 10×buffer 1.25 μl, 25mM MgCl 2 0.8μl, 2.5mM dNTPs 1μl, Taq enzyme (5u / μl) 0.25μl, template DNA 100ng, 20μM primers 0.6μl each, add water to 12.5μl. The PCR reaction system was DNA pre-denatured at 94°C for 5 min, denatured at ...
Embodiment 2
[0038] Example 2: Type AA individuals have stable heredity.
[0039] According to follow-up studies, it has been proved that the parents of AA-type cows have both parents of the A gene (both are AA homozygous; or one is homozygous AA, and the other is AB or AC heterozygous; or both are AA homozygous; Both parties are AB or AC heterozygous), which proves that the AA gene can be inherited figure 2 .
Embodiment 3
[0040] Example 3: Relationship between different genotypes in the regulatory region of Holstein cattle HSP70 gene and heat stress resistance
[0041] 1. Gene frequency and genotype frequency of Holstein cattle heat stress resistance (HSP70 gene regulatory region) locus
[0042] According to the determination method described above in the content of the invention, Holstein cattle blood samples were collected, and the polymorphism of the genetic marker was detected. The results are shown in Table 1.
[0043] Table 1 Genotypes of Holstein Cattle
[0044]
[0045] 2. Changes in rectal temperature, respiratory rate and milk production of cows with different genotypes
[0046] Using SPSS11.5 software, the rectal temperature, respiratory rate and milk production of different genotypes of Holstein cattle were analyzed. The results are shown in Table 2 and Table 3.
[0047] Table 2 Rectal temperature and respiratory rate of each genotype of Holstein cattle
[0048]
[0049]
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