Hmadcha A, De Diego Y, Pintado E
Departamento de Bioquímica Médica y Biología Molecular, Facultad de Medicina y Hospital Universitario Virgen Macarena, Universidad de Sevilla, Spain.
J Lab Clin Med. 1998 Feb;131(2):170-3. doi: 10.1016/s0022-2143(98)90160-3.
The fragile X syndrome is the most frequent form of inherited mental retardation. This is caused by the transcriptional inactivation of the FMR1 gene. The KH domain is an evolutionarily conserved sequence motif present in many RNA-binding proteins including the fragile X mental retardation gene product. We have studied the expression of the gene in fresh leukocytes derived from patients and normal controls by using a reverse transcriptase-polymerase chain reaction (RT-PCR) protocol that amplifies the region of the FMR1 that contains the KH1 and KH2 domains and that has not been used in previous studies. As expected, normal expression was observed in control subjects and carriers, but FMR1 mRNA was absent in male patients with fragile X syndrome. This method was also proved to be useful for testing the expression of FMR1 in samples from several species and tissues. In all cases we obtained a similar and unique transcript. We suggest that RT-PCR from the KH domains could be the method of choice for studying FMR1 expression.
脆性X综合征是遗传性智力迟钝最常见的形式。这是由FMR1基因的转录失活引起的。KH结构域是许多RNA结合蛋白中存在的一种进化保守的序列基序，包括脆性X智力迟钝基因产物。我们使用逆转录酶-聚合酶链反应（RT-PCR）方案研究了该基因在患者和正常对照新鲜白细胞中的表达，该方案扩增了FMR1中包含KH1和KH2结构域的区域，且此前的研究未使用过该区域。正如预期的那样，在对照受试者和携带者中观察到正常表达，但脆性X综合征男性患者中不存在FMR1 mRNA。该方法也被证明对检测来自多个物种和组织的样本中FMR1的表达有用。在所有情况下，我们都获得了相似且独特的转录本。我们认为，从KH结构域进行RT-PCR可能是研究FMR1表达的首选方法。
