2007 Fiscal Year Final Research Report Summary
Studies on development of genome markers and application to colonies in the common marmoset
| Project/Area Number |
17300134
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
KATOH Hideki Hamamatsu University School of Medicine, Experimental Animals Institute, Associate Professor (30142053)
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| Co-Investigator(Kenkyū-buntansha) |
TAKABAYASHI Shuji Hamamatsu University School of Medicine, Experimental Animals Institute, Assistant Professor (70372521)
NISHIKAWA Tetsu Hamamatsu University School of Medicine, Experimental Animals Institute, Assistant Professor (50260584)
TANIOKA Yoshikuni Central Institute for Experimental Animals, Department of Mamoset Research, Researcher (10072406)
SUEMIZU Hiroshi Central Institute for Experimental Animals, Department of Biomedical Research, Researcher (40332209)
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| Project Period (FY) |
2005 – 2007
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| Keywords | Common marmoset / Callithrix jacchus / MHC class II gene / SNP (single nucleotide polvmorphism) / Chromosome / Karvotvpe / Microsatellite DNA marker / Chromosomemap |
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| Research Abstract |
1.Two hundred primer sets were designed to develop microsatellite DNA markers based on the NCBI database archived in 2004. Of 200 markers, 87 showed genetic polymorphisms with five marmosets. Using 42 markers of them selected randomly, different marmoset colonies were studied from a view point of breeding. As a result, differences in the number of alleles and their frequencies were observed one by one. It was revealed that the microsatellite DNA markers are useful for genetic evaluation and breeding of marmoset colonies. Eighty percent of the markers developed in this study were assigned on marmoset chromosomes that are determined with each human chromosome used for DNA probe. Thirteen markers were selected for genetic monitoring which is one of the quality control program. They were amplified using fluorescence-labeled primers. The PCR products were successfully sequenced to determine their sizes at one-base level.2.Exon 2 of the DQA1 and exon 3 of DQB1, which are the marmoset MHC class II genes, were sequenced and SNPs (single nucleotide polymorphisms) were detected in their sequences.3.Isozymes of proteins and enzymes were studied with various electrophoresis techniques and two alleles were observed in each of TRF (transferring) and GPI (glucosephosphate isomerase).4.Red blood cell antigens were studied using 10 antisera for human blood groups (ABO, MN and Rh). Only anti-A antiserum showed positive hemagglutination, but the others showed negative reaction.5.About 1kb D-loop of mitochondrial DNA was sequenced to detect SNPs which may be restriction enzyme recognition sites. As a result, 8 haplotypes were detected using one primer set with five restriction enzymes and another set with one restriction enzyme. G. Preparation of chromosome specimen used for karyotype analysis was successfully done using cultured peripheral lymphocytes and spleen cells.
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