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 /

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.根据2004年存档的NCBI数据库，设计了200组引物来开发微卫星DNA标记。200个标记中，87个显示出与5种狨猴的遗传多态性。利用其中随机选择的42个标记，从育种的角度对不同的狨猴群体进行了研究。结果，等位基因的数量及其频率一一观察到差异。结果表明，微卫星DNA标记可用于狨猴群体的遗传评估和育种。这项研究中开发的标记的 80% 被分配到狨猴染色体上，这些染色体是由用于 DNA 探针的每条人类染色体确定的。选择了13个标记进行遗传监测，这是质量控制项目之一。使用荧光标记的引物对它们进行扩增。 PCR产物成功测序并确定了单碱基水平的大小。2.对狨猴MHC II类基因DQA1的外显子2和DQB1的外显子3进行测序，并检测其序列中的SNP（单核苷酸多态性）。3.通过多种电泳技术对蛋白质和酶的同工酶进行研究，每个中观察到两个等位基因。 TRF（转移）和GPI（磷酸葡萄糖异构酶）。4.使用10种人类血型（ABO、MN和Rh）的抗血清研究红细胞抗原。仅抗A抗血清呈阳性血凝反应，其余均呈阴性反应。5.对线粒体DNA约1kb D-loop进行测序，检测可能为限制性内切酶识别位点的SNP。结果，使用一组具有五种限制性酶的引物和另一组具有一种限制性酶的引物检测到了 8 个单元型。 G.使用培养的外周淋巴细胞和脾细胞成功地制备了用于核型分析的染色体样本。

项目成果

コモンマーモセットのマイクロサテライトマーカーの開発

常见狨猴微卫星标记的开发

• 发表时间: 2005
• 作者: Fujishita;K.;Inoue;K.;Koizumi;S.;高林秀次
• 通讯作者: 高林秀次

Development of microsatellite DNA markers in the common marmoset

普通狨猴微卫星 DNA 标记的开发

• 发表时间: 2005
• 作者: Takabayashi;S
• 通讯作者: S

Genetic polymorphisms of TRF (transferring) and GPI (glucosephosphate isomerase) in the common marmoset

普通狨猴TRF（转移）和GPI（磷酸葡萄糖异构酶）的遗传多态性

• 发表时间: 2008
• 作者: Katoh;H
• 通讯作者: H

マイクロサテライトマーカーを用いたコモンマーモセット(Callithrix jacchus)の多型解析

利用微卫星标记对普通狨猴（Callithrix jacchus）进行多态性分析

• 发表时间: 2005
• 作者: Takabayashi;S;八木橋千枝
• 通讯作者: 八木橋千枝

Anovel Helicobacter species isolated from the feces of the common marmoset (Callithrixjacchus)

从普通狨猴 (Callithrixjacchus) 的粪便中分离出的新型螺杆菌属

• 发表时间: 2007
• 期刊: Fems Microbiology Letters 271
• 作者: Koizumi;S.;Shigemoto-Mogami;Y.;Kaoru;T.;Shinozaki;Y.;Fujishita;K.;Ohsawa;K.;Tsuda;M.;Kohsaka;S. and Inoue;K.;Toung Suk Won
• 通讯作者: Toung Suk Won