SLA typing using the PCR-SSP method and establishment of the SLA homozygote line in pedigreed SNU miniature pigs

摘要

Seoul National University (SNU) miniature pigs represent a closed colony with 24 founder pigs and a well preserved pedigree. Characterization using mRNA sequence analysis was conducted for 6 swine leukocyte antigen (SLA) loci in parental or founder pigs, and 17 defined alleles were detected. Based on these complete coding sequences, 17 sequence specific primers (SSPs) were designed for polymorphic sites. To validate the specificity of each allele SSP, the PCR-SSP was conducted with defined allele clones as templates. PCR-SSP was conducted with the hot start polymerase and touch-down PCR. The parental or found SNU miniature pigs showed overall SLA class I and II heterozygotes. Using the established PCR-SSP method, we conducted SLA typing for breeding stock including 2 pedigreed pigs and identified the novel SLA class II homozygote haplotye (DRA*0201, DRB1*0403, DQA*0102 and DQB1*0701) and 2 SLA homozygote pig lines: SLA class I Hp-3.0 and class II Hp-0.3, and SLA class I Hp-2.0 and class II Hp-0.2. We thought that our PCR-SSP SLA typing method could be applicable for new SLA homozygote line establishment by assignment and scheduled breeding.