Marchetti G, Patracchini P, Gemmati D, DeRosa V, Pinotti M, Rodorigo G, Casonato A, Girolami A, Bernardi F
Centro Studi Biochimici delle Patologie del Genoma Umano, Instituto di Chimica Biologica, Ferrara, Italy.
Hum Genet. 1992 Jul;89(5):497-502. doi: 10.1007/BF00219173.
The 3' portion of the coagulation factor VII gene, containing the activation and serine protease domains, was investigated in four subjects with factor VII deficiency by temperature gradient gel electrophoresis and sequencing of polymerase chain reaction (PCR) products. Molecules displaying an altered melting behaviour were detected in three subjects, and direct sequencing showed two mutations. A G-to-T transversion causing a missense mutation, Cys-310 to Phe, suppresses a disulphide bond conserved in the catalytic domain of all serine proteases. This mutation, which in the homozygous form causes a severe reduction in protease activity (4%), was found in two patients from different Italian regions. A G-to-A transition, which gives rise to a missense mutation, Arg-304 to Gln, and is associated with the factor VII padua variant, was found in the heterozygous form in a subject also affected by von Willebrand disease. Two polymorphic alleles, which differ in one repeat monomer element, were precisely mapped in a region spanning the exon-intron 7 border of the factor VII gene and studied in families with factor VII or X deficiency.
通过温度梯度凝胶电泳和聚合酶链反应（PCR）产物测序，对4名凝血因子VII缺乏症患者凝血因子VII基因的3'部分（包含激活域和丝氨酸蛋白酶域）进行了研究。在3名患者中检测到熔解行为改变的分子，直接测序显示有两个突变。一个由G到T的颠换导致错义突变，即半胱氨酸-310变为苯丙氨酸，破坏了所有丝氨酸蛋白酶催化域中保守的二硫键。这种纯合形式的突变导致蛋白酶活性严重降低（4%），在来自意大利不同地区的两名患者中发现。一个由G到A的转换导致错义突变，即精氨酸-304变为谷氨酰胺，与凝血因子VII帕多瓦变异体相关，在一名同时患有血管性血友病的患者中以杂合形式被发现。两个在一个重复单体元件上不同的多态性等位基因，被精确地定位在凝血因子VII基因第7外显子-内含子边界的一个区域，并在患有凝血因子VII或X缺乏症的家族中进行了研究。
