Cloning, cSNP Identification, and Genotyping of Pig Complement Factor B(CFB) Gene Located on the SLA Class III Region

SLA Class III 영역의 돼지 Complement Factor B(CFB) 유전자의 Cloning, cSNP 동정 및 유전자형 분석

  • Kim, Jae-Hwan (Division of Applied Life Science(BK21 program), Gyeongsang National University) ;
  • Lim, Hyun-Tae (Division of Applied Life Science(BK21 program), Gyeongsang National University) ;
  • Seo, Bo-Yeong (Division of Applied Life Science(BK21 program), Gyeongsang National University) ;
  • Zhong, Tao (Division of Applied Life Science(BK21 program), Gyeongsang National University) ;
  • Yoo, Chae-Kyoung (Division of Applied Life Science(BK21 program), Gyeongsang National University) ;
  • Jung, Eun-Ji (Division of Applied Life Science(BK21 program), Gyeongsang National University) ;
  • Jeon, Jin-Tae (Division of Applied Life Science(BK21 program), Gyeongsang National University)
  • 김재환 (경상대학교 응용생명과학부(BK21 program)) ;
  • 임현태 (경상대학교 응용생명과학부(BK21 program)) ;
  • 서보영 (경상대학교 응용생명과학부(BK21 program)) ;
  • 종타오 (경상대학교 응용생명과학부(BK21 program)) ;
  • 유채경 (경상대학교 응용생명과학부(BK21 program)) ;
  • 정은지 (경상대학교 응용생명과학부(BK21 program)) ;
  • 전진태 (경상대학교 응용생명과학부(BK21 program))
  • Published : 2008.12.31


The primers for RT-PCR and RACE-PCR were designed by aligning the pig genomic sequence and the human complement factor B(CFB) coding sequence(CDS) from the GenBank. Each PCR product was amplified in pig cDNA and sequencing was carried out. The CDS length of pig CFB gene was determined to be 2298 bp. In addition, the pig CDS was more longer than human and mouse orthologs because of insertion and deletion. The identities of porcine nucleotide sequences with those of human and mice were 84% and 80%, and the identities of amino acids were 79% to 77%, respectively. Three complement control protein(CCP) domains, one Von Willebrand factor A(VWFA) domain and a serine protease domain, that are revealed typically in mammals, were found in the pig CFB gene. Based on the CDSs determined, the primers were designed in intron regions for amplification of entire length of exons. In amplification and direct sequencing with genomic DNAs of six pig breeds, three cSNPs(coding single nucleotide polymorphisms) were identified and verified as missense mutations. Using the Multiplex-ARMS method, we genotyped and verified the mutations identified from direct sequencing. To demonstrate recrudescence, we performed both direct sequencing and Multiplex-ARMS with two randomly selected DNA samples. The genotype of each sample exhibited the same results using both methods. Therefore, three cSNPs were identified from pig CFB gene and that can be used for haplotype analysis of the swine leukocyte antigen(SLA) class III region. Moreover, the results indicate that the Multiplex-ARMS method should be powerful for genotyping of genes in the SLA region.



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