Genomic determination of the CR1 (CD35) density polymorphism on erythrocytes using polymerase chain reaction amplification and HindIII restriction enzyme digestion.
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.
Cornillet
P
P
CHU REIMS, Paris, France.
Philbert
F
F
Kazatchkine
M D
MD
Cohen
J H
JH
D001483:Base Sequence; D002874:Chromosome Mapping; D015247:Deoxyribonuclease HindIII; D004587:Electrophoresis, Agar Gel; D004912:Erythrocytes / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011951:Receptors, Complement / Q000235:genetics*; D017463:Receptors, Complement 3b
D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D015247:Deoxyribonuclease HindIII