A kind of genetic marker and its application for evaluating bovine semen quality
A genetic marker and semen technology, applied in the field of genetic markers to evaluate the quality of bovine semen, can solve the problems of no bovine semen kit, no semen quality traits, etc.
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Embodiment 1
[0020] Example 1 Obtaining of bovine FHL2 gene promoter region DNA fragment and establishment of SNP detection method
[0021] The present invention has gone through such as figure 1 The shown process and technical analysis have obtained the bovine FHL2 gene promoter sequence fragment, the fragment length is 1564bp, and its nucleotide sequence is as described in the sequence table SEQ ID NO.1, by performing blast comparison on the above sequence on the NCBI website, in A nucleotide polymorphism (SNP) site is found in the amplified fragment.
[0022] According to bovine FHL2 genome sequence (NCBI reference sequence: NC_037338.1) design following primer pair, concrete sequence is as follows:
[0023] Forward primer 1 (SEQ ID NO.2): 5'-GGGCTTAAAGAACGTGTGTGTG-3',
[0024] Reverse primer 1 (SEQ ID NO.3): 5'-TGACCACCTAGGTCTGGAATGA-3'.
[0025] Forward primer 2 (SEQ ID NO.4): 5'-CATTCCAGACCTAGGTGGTCATT-3',
[0026] Reverse primer 2 (SEQ ID NO.5): 5'-AGGCAAAGCCCTGTATCCAA-3'.
[0...
Embodiment 2
[0036] Example 2: Association analysis of genetic markers of the present invention and bovine semen quality traits
[0037] In order to determine whether the SNP in the promoter region of the bovine FHL2 gene is related to the difference in bovine phenotype, a total of 246 cattle of different breeds (Holstein and Simmental) were selected as test materials. The T879G polymorphic site was genotyped by the first-generation sequencing method, and the specific method was as follows: PCR amplification was performed on the genomic DNA extracted from the semen of 246 cattle with the primer pair shown in SEQ ID NO.4 and SEQ ID NO.5. (Amplification adopts the conditions in Example 1, and the fragment of the amplified product should be the fragment shown in the 648bp-1170bp in SEQID NO.1), after the amplified product is detected by 2% agarose gel electrophoresis, Commissioned Wuhan Tianyi Huiyuan Biotechnology Co., Ltd. for sequencing.
[0038] Three genotypes were detected in this popu...
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