SNP loci for increasing content of fatty acid C14: 0 in chicken and method for breeding high-quality chicken strain
A fatty acid and chicken technology, which is applied in the field of molecular biology, can solve the problems of lack of universality of multi-variety effects, impossibility of livestock breeding practice, and has not yet been carried out, and achieve the effects of high accuracy, low cost, and simple operation
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Embodiment 1
[0038] Example 1. Determination of SNP markers related to fatty acid C14:0 content in chicken
[0039] 1. Genome-wide association analysis (GWAS) obtained SNP molecular markers in the upstream region of FASN gene and chicken fatty acid C14:0 content.
[0040] 1) Experimental animals
[0041]The present invention uses populations from the sixteenth generation chicken IMF selection line (n=256) and control line (n=264), totaling 520 hens. The IMF selection line and the control line are derived from the same basic group (Jingxing Yellow Chicken, Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences). Since 2000, IMF traits have been selected for directional selection. Compared with the control line, the IMF content of the IMF-selected line population was significantly increased (P<0.001), and the shear stress was significantly reduced after selection (ZHAO et al., 2007). During the feeding process, free access to food and water ...
Embodiment 2
[0060] Example 2 The effectiveness and contribution rate of chr18:4910701 and chr18:4911141 mutation-associated fatty acid C14:0 content in chicken meat
[0061] 1. Chr18:4910701 and chr18:4911141 regulate the expression of the associated gene FASN
[0062] 1) Experimental animals
[0063] Using the same animal population as in the GWAS analysis in Example 1 (the sixteenth generation chicken IMF selection line, n=252; and the control line, n=264), the relevant data were used for linkage and variation contribution analysis; Ten mutant individuals and reference individuals carrying the chr18:4910701 site and the chr18:4911141 site were used to detect the expression of the FASN gene.
[0064] 2) The linkage relationship between chr18:4910701 and chr18:4911141 loci
[0065] Use the haplotype linkage analysis method to explore the relationship between the chr18:4910701 site and the chr18:4911141 site, and find that there is a high linkage between the two, such as Figure 4 shown...
Embodiment 3
[0085] Example 3 Establishment of molecular marker detection method for loci chr18:4910701 and chr18:4911141 and their application in breeding
[0086] 1. Establishment of molecular marker detection method
[0087] (1) Primer design
[0088] According to the chicken chromosome 18 DNA sequence provided by the Ensemble website (version 6.0), 2 pairs of specific primers containing these 2 SNP sites were designed using NCBI primer design software. The DNA sequences of the primers are as follows:
[0089] chr18:4910701
[0090] Upstream primer: 5'-TCTTCCCAGGAAGGCGAATTT-3' (SEQ ID NO.1)
[0091] Downstream primer: 5'-AGAATCACGGGATGGTTGGAG-3' (SEQ ID NO.2)
[0092] chr18:4911141
[0093] Upstream primer: 5'-TGGAGGAGGTGAATTCTCACAG-3' (SEQ ID NO.3)
[0094] Downstream primer: 5'-TTTCGGTCGGCAAAAGCG-3' (SEQ ID NO.4)
[0095] (2) PCR reaction program optimization
[0096] The reaction program is: 95°C for 10min, 95°C for 30s, 60°C for 30s, 72°C for 50s, a total of 32 cycles; 72°C ...
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