GENETIC IMPROVEMENT OF JAPANESE BLACK CATTLE BY USING SINGLE LOCUS VNTR MARKERS.

使用单基因座 VNTR 标记对日本黑牛进行遗传改良。

• 批准号: 06454116
• 负责人: TSUJI Soichi
• 金额: $ 4.22万
• 依托单位: KOBE UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454116/
• 关键词: JAPANESE BLACK; VNTRLOSCI; MARKER GENES; REPETITIVE DNA SEQUENCE; MOLECULAR CLONING

Toward efficient improvements of Japanese Black cattle, we made a plan to introduce marker assisted selection procedures. DNA markers are useful tools for the purpose. However, almost all DNA markers now available are dinucleotide repeats. The analysis of dinucleotide erpeats loci requires a special expensive apparatus and high running costs. Therefore, we are planned cloning of tri or tetra nucleotide repeats loci and/or more longer minisatellite loci. Using bovine genomic libray, we had picked up more than 20 clones of tetra nucleotide repeats loci using (GGAT) _4 as a probe. These clones contained (GGAT) _n sequence, but showed incomplete repeats less than 5 repeats, showing no length variation. For (GTG) repeat, we get one clone which is preferable for DNA markers ; heterozygosity of the loci is more than 87% and the polymorphism is easily detectable by using conventional agarose gel electrophoresis. On the other hand, cloning of microsatellite using VNTR 33.15 as a probe, we also obtained 20 positive clones. Employing Southern hybridization technology, we get a few clone of polymorphic.

为了有效地改进日本黑牛，我们制定了一个计划，引入标记辅助选择程序。DNA标记是实现这一目的的有用工具。然而，现在几乎所有可用的DNA标记都是二核苷酸重复序列。二核苷酸重复位点的分析需要昂贵的专用仪器和高昂的运行成本。因此，我们计划克隆三或四核苷酸重复基因座和/或更长的小卫星基因座。利用牛基因组文库，以（GGAT）_4为探针，筛选出20多个四核苷酸重复序列位点的克隆。这些克隆含有（GGAT）_n序列，但重复次数少于5次，无长度变异。对于（GTG）重复，我们得到了一个克隆，这是优选的DNA标记，位点的杂合性大于87%，多态性是很容易检测到的常规琼脂糖凝胶电泳。另一方面，以VNTR 33.15为探针进行微卫星克隆，也获得了20个阳性克隆。利用Southern杂交技术，获得了一些多态性克隆。

项目成果

薫仕ら: "DNAフィンガープリントとアイソザイムパターンによるギンブナクローンの識別" 日本水産学会誌. 62. 747-753 (1996)

Kaorushi 等人：“利用 DNA 指纹和同工酶模式鉴定蠹鱼克隆”，日本水产学会杂志 62. 747-753 (1996)。

Danbaro Gariba et al.,: "Genetic trands from a selection experiment in broiler breeder lines using best linear unbiased prediction methodology." Japanese Poultry Science. 32. 379-386 (1995)

Danbaro Gariba 等人：“使用最佳线性无偏预测方法在肉种鸡品系中进行选择实验的遗传变异。”

費莎ら: "PCR法による食肉・肉製品の肉腫鑑別" 日本畜産学会報. 67. 901-905 (1996)

Kaisa 等人：“通过 PCR 方法鉴定肉和肉制品中的肉瘤”日本动物科学会通报 67. 901-905 (1996)。

Danbaro Gariba et al.,: "Heritbilities and genetic correlations from a selection experiment in broiler breeders using restricted maximum likelyhood." Japanese Poultry Science. 32. 257-266 (1995)

Danbaro Gariba 等人：“使用限制最大似然法对肉种鸡进行选择实验的遗传力和遗传相关性。”

Mannen,H.,: "Identification of sublines of inbred strains of mice and assessment of genetic relationships between sublines or substrains." Experimental Animals. 43. 521-526 (1994)

Mannen，H.，：“小鼠近交系亚系的鉴定以及亚系或亚系之间遗传关系的评估。”