Branchiotai Tibetan pig reproductive performance molecular marker based on ESR gene and primer and screening method of Branchiotai Tibetan pig reproductive performance molecular marker

Abstract

The invention provides an ESR gene-based Branduo Tibetan pig reproductive performance molecular marker and primers thereof, and a method for screening high-reproductive-performance Branduo Tibetan pig individuals by using the marker, the sequence of the molecular marker is a nucleotide sequence as shown in SEQ ID No: 1, the polymorphism of the PvuII site of the ESR gene of the Branduo Tibetan pig is shown as three genotypes of AA, AB and BB, and the BB type is a dominant genotype; a forward primer of a primer sequence of the molecular marker has a nucleotide sequence as shown in SEQ ID No: 2, and a reverse primer of the primer sequence of the molecular marker has a nucleotide sequence as shown in SEQ ID No: 3. The primer can specifically amplify the DNA sequence around the PvuII site of the Tibetan pig ESR gene, the detection efficiency is high, the primer pair is used for molecular marker-assisted breeding, the reproductive performance of the Tibetan pig can be remarkably improved, and large-scale development of the Tibetan pig industry is promoted.

Claims

1. A molecular marker for reproductive performance of Qamdo Tibetan pigs based on the ESR gene, characterized by: The sequence of the molecular marker is a nucleotide sequence shown in SEQ ID No:

1. The polymorphism of the ESR gene PvuII site of the Changdu Tibetan pig is expressed as three genotypes: AA, AB, and BB, among which the BB type is the dominant genotype.

2. The molecular marker for reproductive performance of Qamdo Tibetan pigs based on the ESR gene according to claim 1, characterized in that: The primer sequences of the molecular markers are as follows: Forward primer (F): 5'-CCTGTTTTTACAGTGACTTTTACAGAG-3', the nucleotide sequence shown in SEQ ID No: 2; Reverse primer (R): 5'-CACTTCGAGGGTCAGTCCAATTAG-3', the nucleotide sequence shown in SEQ ID No:

3.

3. A method for screening individuals of Changdu Tibetan pigs with high reproductive performance, characterized in that: The following steps are involved: Step 1, extracting genomic DNA from individual Tibetan pigs; Step 2: using the genomic DNA obtained in step 1 as a template, using a primer pair to perform PCR amplification of the DNA sequence around the PvuII site of the Tibetan pig ESR gene to obtain a molecular marker amplification product; Step 3: Cut the PCR product with PvuII, analyze the product by gel electrophoresis, and determine the genotype based on the number and size of the bands: AA type: 121 bp single band; AB type: two bands of 56bp+65bp; BB type: three bands of 121bp, 56bp, and 65bp; Screen out high-reproductive-performance Tibetan pigs carrying the BB genotype; Wherein, the sequence of the molecular marker is the nucleotide sequence shown in SEQ ID No: 1; The primer pair sequences are as follows: Forward primer (F): 5'-CCTGTTTTTACAGTGACTTTTACAGAG-3', the nucleotide sequence shown in SEQ ID No: 2; Reverse primer (R): 5'-CACTTCGAGGGTCAGTCCAATTAG-3', the nucleotide sequence shown in SEQ ID No:

3.

4. The method for screening high reproductive performance Qamdo Tibetan pig individuals according to claim 3, characterized in that: PCR reaction system 15 μL, the components in the system are: 2×Taq Master Mix: 7 μL, ddH2O: 6 μL, forward primer: 0.5 μL, reverse primer: 0.5 μL, DNA template: 1 μL; PCR reaction conditions included: pre-denaturation: 98°C, 10 minutes; denaturation: 94°C, 50 seconds; annealing: 58°C, 50 seconds; extension: 72°C, 60 seconds / 1 kb; number of cycles: 35 times; final extension: 72°C, 10 minutes, and storage at 4°C.

5. The method for screening high reproductive performance Qamdo Tibetan pig individuals according to claim 3, characterized in that: The enzyme digestion reaction system was as follows: PCR product: 5 μL; ddH2O: 3.5 μL; restriction endonuclease PvuII (10U / μL): 0.5 μL; 10×Buffer: 1 μL; digestion at 37°C for 4 hours.

Images