Identification of DRB alleles in rhesus monkeys using polymerase chain reaction‐sequence‐specific primers (PCR‐SSP) amplification

摘要: Major histocompatibility complex (MHC) class II molecules play a vital role in the regulation of T-cell functions mammalian immune system. Two key features characterize polymorphism MHC haplotypes humans and non-human primates: existence large number alleles, high degree genetic diversity between those alleles. Rhesus monkeys Chimpanzees have been extensively used as relevant models for human diseases transplantation We investigated DRB genes 19 macaques, members 3 families, using polymerase chain reaction with sequence-specific primers (PCR-SSP) denaturing gradient gel electrophoresis (DGGE). After amplification PCR products were purified subjected direct sequencing. Seven animals (Madison #1) typed by DDGE also. report that defined PCR-SSP exhibit concordance data obtained DGGE sequening. Our show prominent variability DRB1 alleles ranging from 1–4 per genotype within these families. This analysis demonstrated most amplicons identical to Mamu-DRB our amplify. However, 98–99% similarity was noticed case Mamu-DRB1*0303, Mamu-DRB6*0103 Mamu-DRB*W201 The observed mismatches located non-polymorphic regions. Thus, family studies rhesus macaques performed molecular methods confirmed multiplicity Mamu-DRB1 haplotype allelic associations published earlier. In addition, we propose more allele (haplotypes): Mamu-DRB1*04-DRB5*03; Mamu-DRB1*04-*DRB*W5; Mamu-DRB1*04*W2. proposed medium-resolution technique appears be highly reproducible discriminatory typing method detecting polymorphisms monkeys.