在狗的杂合突变位点在成纤维细胞mut0 methylmalonic酸血症被聚合酶链反应cDNA克隆。
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Jansen R,莱德利FD
在狗的杂合突变位点在成纤维细胞mut0 methylmalonic酸血症被聚合酶链反应cDNA克隆。
J哼麝猫。1990年11月,47(5):808 - 14所示。
- PubMed ID
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1977311 (在PubMed]
- 文摘
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酶的遗传缺陷methylmalonyl有机酸代谢的辅酶a变位酶引起障碍称为“傻瓜methylmalonic酸血症”。Various phenotypes of mut methylmalonic acidemia are distinguished by the presence (mut-) or absence (mut0) of residual enzyme activity. The recent cloning and sequencing of a cDNA for human methylmalonyl CoA mutase enables molecular characterization of mutations underlying mut phenotypes. We identified compound heterozygous mutations in a mut0 fibroblast cell (MAS) line by cloning the methylmalonyl CoA mutase cDNA by using the polymerase chain reaction (PCR), sequencing with internal primers, and confirming the pathogenicity of observed mutations by DNA-mediated gene transfer. Both mutations alter amino acids common to the normal human, mouse, and Propionibacterium shermanii enzymes. This analysis points to evolutionarily preserved determinants critical for enzyme structure or function. The application and limitation of cDNA cloning by PCR for the identification of mutations are discussed.