Identification of a Gly862 to Ser substitution in the type I collagen gene from a single spermatozoon.
Individual spermatozoa from the father of two affected infants with osteogenesis imperfecta were separated by dilution and micromanipulation. A segment of the type I collagen gene containing the mutant region was amplified by nested polymerase chain reaction and sequenced. Among 40 individual spermatozoa, 15 specimens were identified as mutants with a substitution of guanine3208 to adenine (glycine862 to serine) while 18 and seven specimens were of the wild and mixed types respectively. Through this study, we have established a molecular procedure that can be considered as prerequisite for preimplantation diagnosis of genetic disorders with a single point mutation.
Iida
T
T
Department of Obstetrics and Gynecology, Nagoya City University Medical School, Japan.
Suzumori
K
K
Ikuta
K
K
Tanemura
M
M
Yagami
Y
Y
Okamoto
T
T
Hata
A
A
D003094:Collagen / Q000235:genetics*; D004247:DNA / Q000302:isolation & purification; D017931:DNA Primers; D005260:Female; D005307:Fertilization in Vitro; D005820:Genetic Testing; D005998:Glycine; D006801:Humans; D008297:Male; D009030:Mosaicism / Q000235:genetics; D010013:Osteogenesis Imperfecta / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D017422:Sequence Analysis, DNA; D012694:Serine; D013094:Spermatozoa / Q000737:chemistry*
D017931:DNA Primers; D012694:Serine; D003094:Collagen; D004247:DNA; D005998:Glycine