Detection of a genetic mutation for myotonia congenita among Miniature Schnauzers and identification of a common carrier ancestor
Detection of a genetic mutation for myotonia congenita among Miniature Schnauzers and identification of a common carrier ancestor
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DOI：
10.2460/ajvr.2002.63.1443
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发表时间：
2002-10-01
影响因子：
1
通讯作者：
Giger, U
中科院分区：
文献类型：
--
作者：
Bhalerao, DR;Rajpurohit, Y;Giger, U
2.建立一种分子遗传学检测方法，检测导致小型雪纳瑞犬先天性肌强直的突变型骨骼肌氯离子通道（CIC-1）等位基因，并分析患病犬与携带者犬的关系。方法-通过使用一对独特的引物扩增CIC-1等位基因突变周围的序列。用限制性内切酶Hpy CH 4 III消化聚合酶链反应（PCR）产物，并在6%聚丙烯酰胺凝胶上分离。从所有可用的载体和受影响的狗的谱系进行了分析，并建立了一个复合pedigree.Results-enzyme消化正常CIC-1等位基因的PCR产物导致3个片段的175，135，和30 bp，而突变等位基因的PCR产物导致的片段只有175和165 bp。在372只小型雪纳瑞犬中，292只（78.5%）正常，76只（20.4%）携带者，4只（1.1%）受影响（肌强直）犬。突变等位基因频率为0.113。系谱分析显示，一个流行的父亲，记录是一个载体，是一个共同的祖先，所有的运营商和受影响的dogs.Conclusions和临床相关性，一个PCR为基础的酶消化DNA测试开发。这种疾病的突变等位基因在小型雪纳瑞中很常见，它们与共同的携带者祖先有关。繁殖狗应该通过这种特定的DNA测试，以帮助限制这种有害突变的传播。
Objective-To develop a molecular genetic test to detect the mutant skeletal muscle chloride channel (CIC-1) allele that causes myotonia congenita in Miniature Schnauzers and to analyze the relationship of affected and carrier dogs.Animals-372 Miniature Schnauzers from the United States, Canada, Australia, and Europe that were tested between March 2000 and October 2001.Procedure-The sequence surrounding the mutation in the CIC-1 allele was amplified by use of a unique pair of primers. Polymerase chain reaction (PCR) products were digested with the restriction enzyme Hpy CH4 III and separated on a 6% polyacrylamide gel. Pedigrees from all available carrier and affected dogs were analyzed, and a composite pedigree was established.Results-Enzyme digestion of PCR products of the normal CIC-1 allele resulted in 3 fragments of 175, 135, and 30 bp, whereas PCR products of the mutant allele resulted in fragments of only 175 and 165 bp. Of the 372 Miniature Schnauzers, 292 (78.5%) were normal, 76 (20.4%) were carriers, and 4 (1.1%) were affected (myotonic) dogs. Frequency of the mutant allele was 0.113. Pedigree analysis revealed that a popular sire, documented to be a carrier, was a common ancestor of all carriers and affected dogs.Conclusions and Clinical Relevance-A PCR-based enzyme digestion DNA test was developed. The mutant allele for this disease is frequent in Miniature Schnauzers that are related to a common carrier ancestor. Breeding dogs should be tested by this specific DNA test to help limit the spread of this deleterious mutation.