Hong C M, Ohashi T, Yu X J, Weiler S, Barranger J A
Section of Molecular Genetics, University of Southern California, School of Medicine, Childrens Hospital, Los Angeles 90027.
DNA Cell Biol. 1990 May;9(4):233-41. doi: 10.1089/dna.1990.9.233.
Enzymatically amplified RNA transcripts were used to analyze the full coding region of the glucocerebrosidase gene from Gaucher disease patients. Two previously undescribed mutations were identified. One mutation consists of a single-base substitution in three different codons: codon 444, Leu (CTG) to Pro (CCG); codon 456, Ala (GCT) to Pro (CCT); and codon 460, Val (GTG) to Val (GTC). This mutant is called "pseudo pattern" (psi) because it is identical in sequence to a small region of the pseudogene in exon 10 (Horowitz et al., 1989). The other new mutation is a single-base substitution (C to T) resulting in the substitution of Cys for Arg in codon 463. These mutations in the human gene were duplicated in wild-type cDNA and expressed in 3T3 cells. The human mutant proteins were isolated by immunoaffinity and shown to have altered enzymatic properties demonstrating the causality of these two allelic mutations for Gaucher disease.
酶促扩增的RNA转录本用于分析戈谢病患者葡糖脑苷脂酶基因的完整编码区。鉴定出两个先前未描述的突变。一个突变由三个不同密码子中的单碱基替换组成:密码子444,亮氨酸(CTG)突变为脯氨酸(CCG);密码子456,丙氨酸(GCT)突变为脯氨酸(CCT);密码子460,缬氨酸(GTG)突变为缬氨酸(GTC)。这种突变体被称为“假模式”(psi),因为它在序列上与外显子10中假基因的一个小区域相同(霍洛维茨等人,1989年)。另一个新突变是单碱基替换(C突变为T),导致密码子463中的半胱氨酸替换精氨酸。人类基因中的这些突变在野生型cDNA中复制并在3T3细胞中表达。通过免疫亲和法分离出人类突变蛋白,并显示其酶学性质发生改变,证明了这两个等位基因突变与戈谢病的因果关系。