Rapid assignment of the swine major histocompatibility complex (<i>SLA</i>) class I and II genotypes in Clawn miniature swine using PCR‐SSP and PCR‐RFLP methods

• Asako Ando

• Masao Ota

• Masaharu Sada

• Yoshihiko Katsuyama

• Rieko Goto

• Atsuko Shigenari

• Hisako Kawata

• Tatsuya Anzai

• Takahiro Iwanaga

• Yukari Miyoshi

• Nobuyuki Fujimura

• Hidetoshi Inoko

抄録

Abstract:  Background:  Inbred miniature swine with defined novel SLA haplotypes will be useful in allo‐ and xeno‐transplantation studies, which can be carried out representing variable combinations of SLA haplotypes. Methods:  In Clawn miniature swine, two haplotypes ( c1 and c2 ) and one crossover haplotype ( c3 ) have been assigned by nucleotide sequence determination of RT‐PCR products of the three SLA classical class I genes and two SLA class II genes. To select SLA class I and II homozygotes in Clawn miniature swine individuals, we developed a rapid and simple SLA‐class I‐ and II‐DNA typing method by a combination of polymerase chain reaction‐sequence specific primer (PCR‐SSP) and polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) techniques. Results:  Seven allele specific primer pairs were designed for amplification of the second exons of three SLA class I genes, SLA‐1 , SLA‐2 , and SLA‐3 , and one SLA class II gene, DRB1 . Furthermore, based on PCR‐RFLP patterns in the SLA‐DQB1 gene, two allelic variants were recognized in the second exon in the Clawn miniature swine. Three haplotypes, c1 , c2 and c3 , were simply identified by the combination of PCR‐SSP and PCR‐RFLP methods in 22 samples from five families. A single allele at each of the class I and II genes was also observed in seven samples as SLA class I and II homozygotes with either the c1 or c2 haplotype. Conclusions:  The combination of PCR‐SSP and PCR‐RFLP methods facilitate the rapid identification of the three haplotypes and SLA class I and II homozygotes in individual Clawn miniature swine.

収録刊行物

• Xenotransplantation

  Xenotransplantation 12 (2), 121-126, 2005-02

  Wiley

キーワード

• Transplantation
• Immunology

詳細情報

• CRID: 1361137043556250880
• NII論文ID: 30013468794
• DOI: 10.1111/j.1399-3089.2005.00204.x
• ISSN: 13993089
• ISSN: 0908665X
• Web Site: https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1399-3089.2005.00204.x
• Web Site: https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1399-3089.2005.00204.x
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