Rapid detection of polymorphisms of the nitric oxide cascade.
NOS3 (endothelial nitric oxide (NO) synthase) and p22phox (subunit of NAD(P)H oxidase) are two genes whose products are involved in formation and degradation of NO, a ubiquitous signaling molecule largely responsible for the maintenance of normal endothelial function. The G894T polymorphism of NOS3 and the C242T polymorphism of p22phox are reportedly associated with numerous cardiovascular diseases. For each polymorphism we developed a rapid and reliable method with the hybridization probes format on the LightCycler and compared it with conventional PCR-restriction fragment length polymorphism (PCR-RFLP) analysis with regard to reliability, duration and cost. The new methods are more reliable, faster and less expensive than PCR-RFLP analysis and therefore represent a significant advantage in the detection of two candidate risk factors for cardiovascular diseases.
Weiss
Johanna
J
Department of Internal Medicine VI, Clinical Pharmacology and Pharmacoepidemiology, University of Heidelberg, Germany. johanna_weiss@med.uni-heidelberg.de
Fricker
Ruth
R
Haefeli
Walter E
WE
D000483:Alleles; D001483:Base Sequence; D002610:Cheek; D004735:Energy Transfer; D005453:Fluorescence; D005838:Genotype; D006358:Hot Temperature; D006801:Humans; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D009252:NADPH Dehydrogenase / Q000097:blood / Q000235:genetics*; D019255:NADPH Oxidases; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000097:blood / Q000235:genetics*; D052250:Nitric Oxide Synthase Type III; D009691:Nucleic Acid Denaturation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010750:Phosphoproteins / Q000097:blood / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic / Q000235:genetics; D017422:Sequence Analysis, DNA
D026901:Membrane Transport Proteins; D009838:Oligodeoxyribonucleotides; D010750:Phosphoproteins; D009569:Nitric Oxide; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D019255:NADPH Oxidases; C086986:CYBA protein, human; D009252:NADPH Dehydrogenase