Biery B J, Stein D E, Morton D H, Goodman S I
Department of Cellular and Structural Biology, University of Colorado School of Medicine, Denver 80262, USA.
Am J Hum Genet. 1996 Nov;59(5):1006-11.
The structure of the human glutaryl coenzyme A dehydrogenase (GCD) gene was determined to contain 11 exons and to span approximately 7 kb. Fibroblast DNA from 64 unrelated glutaric acidemia type I (GA1) patients was screened for mutations by PCR amplification and analysis of SSCP. Fragments with altered electrophoretic mobility were subcloned and sequenced to detect mutations that caused GA1. This report describes the structure of the GCD gene, as well as point mutations and polymorphisms found in 7 of its 11 exons. Several mutations were found in more than one patient, but no one prevalent mutation was detected in the general population. As expected from pedigree analysis, a single mutant allele causes GA1 in the Old Order Amish of Lancaster County, Pennsylvania. Several mutations have been expressed in Escherichia coli, and all produce diminished enzyme activity. Reduced activity in GCD encoded by the A421V mutation in the Amish may be due to impaired association of enzyme subunits.
已确定人类戊二酰辅酶A脱氢酶（GCD）基因的结构包含11个外显子，跨度约为7 kb。通过PCR扩增和SSCP分析，对来自64名无关的I型戊二酸血症（GA1）患者的成纤维细胞DNA进行突变筛查。将电泳迁移率改变的片段亚克隆并测序，以检测导致GA1的突变。本报告描述了GCD基因的结构，以及在其11个外显子中的7个中发现的点突变和多态性。在不止一名患者中发现了几种突变，但在普通人群中未检测到一个普遍存在的突变。正如系谱分析所预期的那样，单个突变等位基因在宾夕法尼亚州兰开斯特县的老派阿米什人中导致GA1。几种突变已在大肠杆菌中表达，并且都产生了降低的酶活性。阿米什人中由A421V突变编码的GCD活性降低可能是由于酶亚基的缔合受损。
