Study Of DNA Technology For Porcine Meat Traceability Based On SNPs Markers

In order to explore the establishment of traceablity identity system for pork safety, this study built DNA fingerprinting on pigs by using the third generation genetic marker—single nucleotide polymorphisms(SNPs). Based on the difference of individual DNA fingerprinting we could identify each animal. Once the accidents of pork safety happened, the system could traced the source of individuals quickly by markers. This paper aimed to find the effective SNP locis, and establish the individual DNA fingerprinting—SNP panels.The candidate SNP locis could be selected from a database, or the unknown new SNP could be discovered, we used PCR-RFLP method to genetype for the candidate SNPs. Assess and analysis the polymorphism of candidate SNP locis by calculating the allele frequency and heterozygosity of each locus. We selected the sites which have content-rich polymorphic information as the individual valid markers. The sequencing method could be used to discover the new SNP, and then the effectiveness was test. Finally, the effective SNP locis formed a SNP panel. Then the effectiveness of this panel would be test for individual indentification through establishing simulated traceability system of individual identification. The main results of this paper are as follows:1. In this study, we selected 23 candidates SNP from a large number of literatures, and found that 11 SNP marker locis meet the standerd that the distribution of allele gene frequency was balance, the heterozygosity was high and the polymorphism information was rich on majority of breeds. They could be used as the valid markers.2. Many conserved DNA sequences through NCBI database was to be obtained for excavating the unknown new SNP. The primers were designed using the software Primer 5.0, and then the annealing temperature of PCR were groped. After the pre-experiment., we tested the efficiency of each primer and choose the high efficiency primers to amplificate. The amplified products were sequenced and compared, then the new SNP sites would be found. Four SNP which could be analysised by PCR-RFLP were discovered. After genotyping and SNP evaluation, we ultimately found 3 new SNP that are valid markers.3. Thirteen effective sites from candidate SNP were selected to form a SNP panel. Three hundred and ten pigs samples were markered (the species was unknown) randomly from Shanghai slaughter to establish a simulated recognition system for pork traceablity. The recognition results were fully identified.