DNA extraction from bovine mummified fetuses and detection of factor XI gene deficiency in the mummies.
DNA extraction from bovine mummified fetuses and detection of factor XI gene deficiency in the mummies.
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牛木乃伊胎儿 DNA 提取及木乃伊因子 XI 基因缺陷检测。
DOI：
10.1262/jrd.16080
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
M. Moriyoshi
中科院分区：
文献类型：
--
作者：
M. Ghanem;M. Nishibori;T. Nakao;M. Moriyoshi
从牛木乃伊组织中提取的基因组DNA是检测可能导致胎儿异常的一些基因的宝贵材料。在这项研究中，牛基因组DNA是从10个牛木乃伊胎儿的硬化组织样本中提取的。用苯酚/氯仿-乙醇法从2 g木乃伊组织中提取的基因组DNA量很低（小于4微克/毫升）。将提取的DNA用GenomiPhi DNA扩增系统进行扩增。扩增后，所有样品的DNA量均增加到100 μ g/ml以上。该扩增系统被证明是扩增木乃伊胎儿基因组DNA的好工具。扩增的基因组DNA用于检测木乃伊的因子XI基因缺陷，这是一种常染色体隐性缺陷，涉及内在血液凝固途径的早期阶段。用PCR扩增了木乃伊因子XI基因的第12外显子。十个木乃伊胎儿中有两个是因子XI基因的杂合，这表明存在两个扩增的DNA片段，分别为320 bp和244 bp。荷斯坦牛已经描述过因子XI缺乏症。然而，没有关于牛胎儿的报道。本研究从牛胎儿木乃伊中提取并扩增了DNA，并成功地对样品进行了因子XI基因缺乏的测试。
Genomic DNA extracted from bovine mummified tissue is valuable material for detection of some genes that may contribute to fetal abnormalities. In this study bovine genomic DNA was extracted from the hardened tissue samples of ten bovine mummified fetuses. The amount of genomic DNA extracted from 2 g of the mummified tissues by the phenol/chloroform-ethanol method was low (less than 4 microg/ml) for all samples. The extracted DNA was then amplified by the GenomiPhi DNA amplification system. After amplification, the amount of DNA was increased to more than 100 microg/ml for all samples. This amplification system was shown to be a good tool for amplifying the genomic DNA of the mummified fetuses. The amplified genomic DNA was used for testing the mummies for Factor XI gene deficiency, an autosomal recessive deficiency involved in the early stages of the intrinsic blood coagulation pathway. Exon 12 of the Factor XI gene of the mummies was amplified by PCR. Two of the ten mummified fetuses were heterozygous for the Factor XI gene as indicated by the presence of two amplified DNA fragments of 320 bp and 244 bp. Factor XI deficiency has already been described in Holstein cattle. However, no report is available for bovine fetus. In this study, DNA was extracted and amplified from the bovine mummified fetuses, and the samples were successfully tested for Factor XI gene deficiency in the mummies.