Cornillet P, Philbert F, Kazatchkine M D, Cohen J H
CHU REIMS, Paris, France.
J Immunol Methods. 1991 Feb 15;136(2):193-7. doi: 10.1016/0022-1759(91)90006-2.
The density of CR1 (the C3b receptor, CD35) on erythrocytes from normal individuals is determined by a codominant bi-allelic system associated with a single base mutation within an intron of the CR1 structural gene, leading to an additional polymorphic HindIII endonuclease site. The CR1 genotype is determined by HindIII digestion of genomic DNA and Southern blotting. We have developed a method based on polymerase chain reaction (PCR) amplification of the genomic DNA fragment of interest followed by HindIII endonuclease digestion and agarose gel electrophoresis which permits a more rapid and reliable determination of the CR1 genotype. The method is suitable for large scale clinical studies in diseases with altered expression of CR1 on erythrocytes.
正常个体红细胞上CR1(C3b受体,CD35)的密度由一个共显性双等位基因系统决定,该系统与CR1结构基因内含子内的单个碱基突变相关,导致一个额外的多态性HindIII核酸内切酶位点。CR1基因型通过基因组DNA的HindIII消化和Southern印迹法确定。我们开发了一种基于聚合酶链反应(PCR)扩增感兴趣的基因组DNA片段,随后进行HindIII核酸内切酶消化和琼脂糖凝胶电泳的方法,该方法能够更快速、可靠地确定CR1基因型。该方法适用于对红细胞上CR1表达改变的疾病进行大规模临床研究。