Studies on development of genome markers and application to colonies in the common marmoset

狨猴基因组标记的开发及其在群体中的应用研究

• 批准号: 17300134
• 负责人: KATOH Hideki
• 金额: $ 4.88万
• 依托单位: Hamamatsu University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17300134/
• 关键词: Common marmoset; Callithrix jacchus; MHC class II gene; SNP (single nucleotide polvmorphism); Chromosome; Karvotvpe; Microsatellite DNA marker; Chromosomemap; DQA1遣伝子; Microsatellite marker; 実験動物; 高等霊長類; ミトコンドリアDNA多型; PCR-RFLP; Transferrin /; Common marmoset; Callithrix jacchus; MHC class II gene; SNP (single nucleotide polvmorphism); Chromosome; Karvotvpe; Microsatellite DNA marker; Chromosomemap; DQA1遣伝子; Microsatellite marker; 実験動物; 高等霊長類; ミトコンドリアDNA多型; PCR-RFLP; Transferrin /

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.

1.设计了两百个底漆集，以基于2004年存档的NCBI数据库开发微卫星DNA标记。在200个标记中，有87个显示出具有五根质子的遗传多态性。使用42个标记随机选择，从繁殖的角度研究了不同的摩尔群落菌落。结果，一一观察到等位基因及其频率的差异。据揭示，微卫星DNA标记对于果泥群菌落的遗传评估和繁殖很有用。在这项研究中开发的标记中有80％是在用用于DNA探针的每个人类染色体确定的Marmoset染色体上分配的。选择了13个标记进行基因监测，这是质量控制计划之一。使用荧光标记的引物扩增它们。将PCR产物成功测序以确定其在一个基水平上的尺寸。2.DQB1的DQA1和外显子3的EXON 2（它们是Marmoset MHC MHC II类基因）进行了测序，并在其序列中检测到了蛋白质和两种蛋白质，并在其序列中检测到SNP（单核苷酸多态性）。 TRF（转移）和GPI（葡萄糖磷酸异构酶）的of。4。使用10抗血细胞抗原对人类血型（ABO，MN和RH）进行了研究。只有抗A抗血清显示阳性血凝，但其他抗血清显示为阴性反应。5.1.Abbob 1kb D-loop的线粒体DNA测序以检测可能是限制酶识别位点的SNP。结果，使用一个具有五种限制酶的引物和一种限制酶的一个引物，检测到8个单倍型。 G.使用培养的外周淋巴细胞和脾细胞成功进行了用于核型分析的染色体标本。