Uchihi R

Analysis of allelic structures at the D7S21 (MS31A) locus in the Japanese, using minisatellite variant repeat mapping by PCR (MVR-PCR).

To sample the diversity of allelic structures at the D7S21 (MS31A) locus in the Japanese, allele‐specific minisatellite variant repeat mapping using polymerase chain reaction (MVR‐PCR) was performed on genomic DNA from a number of Japanese individuals. Three polymorphic positions in the MS31A 5′ flanking DNA were typed from 214 un‐related Japanese, and the distribution of haplotypes was analysed. Allele‐specific MVR‐PCR, using primers that discriminate between different alleles at these polymorphic positions in heterozygous individuals, allows single alleles to be mapped from genomic DNA in approximately 80% of Japanese. 149 Japanese alleles have been mapped to date and all of them, except for two pairs of indistinguishable alleles, have different internal structures. More than half of the mapped alleles showed similar regions of internal structure to other alleles and were classified into groups on this basis.

Major histocompatibility complex DQA1 nucleotide sequences of gelada baboon (Theropithecus gelada), olive baboon (Papio anubis), and yellow baboon (Papio cynocephalus)

Non-human primates provide valuable models to investigate the causes of human diseases, to test candidate vaccines against infectious pathogens, and to evaluate the efficacy of fertility regulating agents. Understanding the characteristics of the primate major histocompatibility complex (MHC) may be critical in the interpretation of results of vaccine trials and investigations of possible associations between MHC and disease. Routine typing of primate MHC alleles has been hampered by the unavailability of specific immunological reagents and allele-specific oligonucleotide probes. Hence, there is a need to generate adequate nucleotide sequence information that can be used to design polymerase chain reaction (PCR)based techniques for the typing of primate MHC alleles. Among the primates, MHC-DQA1 of gorilla, chimpanzee, and rhesus macaques have been well characterized (Christ et al. 1994; Sauermann et al. 1995; Slierendregt et al. 1993; Kenter et al. 1993; Bontrop et al. 1995). In the present study, genomic DNA was isolated from ten baboons(Papio anubisN = 5,Papio cynocephalus N = 2, Theropithecus gelada N = 2, Papio hamadryasN = 1) and the polymorphic second exon of the DQA1 locus was amplified using GH26 and GH27 PCR primers (Scharf et

Nucleotide sequences of the major histocompatibility complex DQA1 locus of Cercopithecus monkeys

Major histocompatibility complex (MHC) gene products are highly polymorphic. This characteristic contributes to antigen binding repertoire and thus plays a vital role in the generation of effective immune responses to a variety of infectious pathogens (Bontrop et al. 1996; Parham and Ohta 1996). The nucleotide sequence diversity of the polymorphic MHC class II genes is located in the second exon and encodes amino acid residues that are found in the antigen binding site (Slierendregt et al. 1993). Previous studies have shown that most Old World monkeys lack MHC-DQA2andDQB2 (Bontrop et al. 1995; Kenter et al. 1992). In contrast, primate DQA1 is polymorphic and some allelic lineages are shared by humans, apes, and monkeys (Bontrop et al. 1995; Sauermann et al. 1995; Christ et al. 1994). In the present study, we analyzed the DQA1 locus of twenty-two AfricanCercopithecusmonkeys maintained at the Institute of Primate Research, Nairobi, Kenya. These primates included three African green monkeys (C. aethiops),seventeen Sykes monkeys (C. mitis)representing the three Kenyan subspecies, including five highland sykes (C. m. kolbi),seven lowland sykes (C. m. albogularis) , five blue sykes(C. m. stuhlmanni) , and two De-Brazza’s monkeys(C. neglectus ). In addition, two black-and-white colobus monkeys(Colobus guereza) and one black mangabey (Cercocebus aterrimus) were analyzed. Genomic DNA was isolated from peripheral blood lymphocytes and the DQA1 gene region was amplified using GH26 and GH27 polymerase chain reaction (PCR) primers (Scharf et al. 1986). The PCR products (242 or 239-bp) were cloned using the TA cloning kit (Invitrogen, San Diego, CA) and nucleotide sequences determined using an automated DSQ-1 sequencer (Shimadzu, Japan) and cycle sequencing reagents (Perkin-Elmer, Branchburg, NJ). For each individual, sequences were determined from at least two PCR amplifications and a minimum of three plasmid clones. Computerassisted analysis of the obtained nucleotide sequences was performed to compare similarity between individuals of the same and different species and also with published human and primate sequences using DNA database (DDBJ) FASTA (version 1.30). The designation of the genes represented in these sequences has been standardized according to the recommendation contained in an earlier proposal (Klein et al. 1990). An alignment of representative sequences from each species is shown in Fig. 1. In some instances, the sequences derived from different individuals were identical ( 499%) and one representative was used in the alignment. The results of this study provide additional sequence information on fifteen new MHC-DQA1alleles of African Old World monkeys and extends the previous compilation of non-human primate DQA1sequences (Bontrop 1994). The nucleotide sequence data reported in this paper have been submitted to the EMBL/GeneBank databases and have been asigned the following accession numbers: D88582 (African green monkey, VER 1548), D88583 (African green monkey, VER 1621), D88739 (African green monkey, GRIV 14), D88675 (blue sykes, BLU 26), D88676 (blue sykes, BLU 29), D88677 (Blue sykes, BLU 28), D89536 (blue sykes, BLU 30), D89537 BLU 31), D88678 (highland sykes, SYK 269), D88679 (highland sykes, SYK 276), D88680 (highland sykes, SYK 354), D89531 (highland sykes, SYK 191), D89532 (highland sykes, SYK 314), D88681 (lowland sykes monkey, SYL 51), D88682 (lowland sykes monkey, SYL 70), D88683 (lowland sykes monkey, SYL 66), D88684 (lowland sykes monkey, SYL 120), D889534 (lowland sykes, SYL 73), D89534 (lowland sykes, SYL 1 19), D89535 (lowland sykes, SYL 117), D88580 (De Brazza’s monkey, DEB 53), D88581 (De Brazza’s monkey, DEB 54), D88579 (black mangabey, BLM 29) and D88578 (black and white colobus monkey, COL 38), and D89530 (black and white colobus monkey, COL 32)