Almeida Andresa Cardoso Grandini, Pasqualim Gabriela, Mayer Fabiana Q, Schwartz Ida Vanessa Doderlein, Souza Carolina F, Giugliani Roberto, Matte Ursula
Gene Therapy Center, Research Center, Hospital de Clínicas de Porto Alegre, Porto Alegre, RS, Brazil.
Diagn Mol Pathol. 2012 Mar;21(1):53-5. doi: 10.1097/PDM.0b013e318230f021.
Nonsense-mediated decay (NMD) is a mechanism of the recognition and degradation of messenger RNA containing a premature stop codon. Nonsense mutations are the main mutations that lead to Mucopolysaccharidosis type I. To determine the effect of NMD on correct genotyping based on cDNA sequencing, we standardized the sequencing from alpha-L-iduronidase gene cDNA molecules and validated this process for a group of patients whose mutations had been previously identified by DNA analysis. Although the whole gene could be amplified in 5 polymerase chain reactions, cDNA proved unsuitable for molecular analysis as patients bearing splice site and nonsense mutations were not genotyped.
无义介导的衰变（NMD）是一种识别和降解含有提前终止密码子的信使核糖核酸的机制。无义突变是导致I型黏多糖贮积症的主要突变。为了确定NMD对基于互补脱氧核糖核酸（cDNA）测序的正确基因分型的影响，我们对α-L-艾杜糖醛酸酶基因cDNA分子的测序进行了标准化，并针对一组先前通过DNA分析鉴定出突变的患者验证了这一过程。尽管整个基因可以在5次聚合酶链反应中扩增，但cDNA被证明不适用于分子分析，因为携带剪接位点和无义突变的患者无法进行基因分型。
