Levy N J, Ramesh N, Cicardi M, Harrison R A, Davis A E
Department of Pediatrics, Harvard Medical School, Boston, MA 02115.
Proc Natl Acad Sci U S A. 1990 Jan;87(1):265-8. doi: 10.1073/pnas.87.1.265.
Identical single-base changes in the C1 inhibitor gene that may result in dysfunctional inhibitor proteins are described in two different families with type II hereditary angioneurotic edema. Initially, a restriction fragment length polymorphism was defined that resulted from loss of a Pst I site within exon VIII, which encodes the region containing the reactive center. Exon VIII from the normal and abnormal allelles was amplified by the polymerase chain reaction. Amplified DNA product was cloned into plasmid pUC18; clones representing normal and mutant allelles were distinguished by the presence and absence, respectively, of the Pst I restriction site. DNA sequence analysis revealed a G----A mutation in the codon for alanine-436, which would result in replacement with a threonine residue. This position is nine amino acid residues amino-terminal to the reactive-center arginylthreonine peptide bond. In contrast, previously defined mutations in type II hereditary angioneurotic edema result in replacement of the reactive-center arginine.
在两个患有II型遗传性血管性水肿的不同家族中,描述了C1抑制剂基因中相同的单碱基变化,这些变化可能导致功能失调的抑制剂蛋白。最初,定义了一种限制性片段长度多态性,其由外显子VIII内Pst I位点的缺失引起,该外显子编码包含反应中心的区域。通过聚合酶链反应扩增正常和异常等位基因的外显子VIII。将扩增的DNA产物克隆到质粒pUC18中;分别通过Pst I限制性位点的存在和缺失来区分代表正常和突变等位基因的克隆。DNA序列分析显示丙氨酸-436密码子发生G----A突变,这将导致被苏氨酸残基取代。该位置在反应中心精氨酰苏氨酸肽键的氨基末端九个氨基酸残基处。相比之下,先前定义的II型遗传性血管性水肿突变会导致反应中心精氨酸的取代。