Clone and application of molecule mark used as swine mark auxiliary selection and correlated with immunity
A marker-assisted selection and molecular marker technology, applied in the field of livestock genetic engineering, can solve problems such as the function of pig BCL10 gene that has not been fully studied.
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Embodiment 1
[0034] (1) BCL10 gene CDS cloning and partial DNA sequence amplification
[0035] (1) Primer design
[0036] Use human BCL10 gene mRNA (GenBank accession number: NM 003921) as an information probe, utilize the BLAST tool in NCBI to do homologous sequence screening in the GenBank pig EST database, and obtain a series of ESTs with more than 90% homology (fragment length greater than 100bp), use ENTREZ (http: / / www.ncbi.nlm.nih.gov / Web / Search / index.html) to query the corresponding sequences of these ESTs in NCBI, and then use the sequence analysis software DNAStar The SeqMan program constructs porcine EST-contigs. According to the homologous comparison with the human BCL10 gene mRNA, the start and stop codon positions were preliminarily determined, and primers were designed to clone the cDNA sequence (including the full-length CDS and part of the 3'-UTR) as shown in SEQ ID NO: 1 in the sequence table According to the result of comparison with the full-length DNA sequence of the ...
Embodiment 2
[0062] The experimental pigs came from the Landrace pigs of the Shuitai original pig farm of Guangdong Huanong Wenshi Animal Husbandry Co., Ltd. (the blood of foreign pig breeds). The parents were 17 Landrace boars and 36 Landrace sows. The first piglets, parents and offspring were all genotype tested. The offspring collected anticoagulant and non-anticoagulant blood at 0, 17, and 32 days of age, and routine blood routine and antibody levels were detected by conventional methods.
[0063] The traits analyzed are mainly immune traits, including porcine reproductive and respiratory syndrome (PRRS) virus antibody levels, classical swine fever virus antibody levels, pseudorabies antibody levels and 18 blood routine items. According to the population structure of the collected samples, the applicant used a mixed model to statistically analyze the genotype effect of the SNPs of the BCL10 gene and its relationship with immune indicators:
[0064] Y=Xβ+Zγ+e
[0065] Among them, Y is ...
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