Identification of human TGF-beta1 signal (leader) sequence polymorphisms by PCR-RFLP.
Links between disease susceptibility and genetically determined variation in human cytokine expression have recently been described. This has led to a demand for simple methods of identifying cytokine gene polymorphisms of potential clinical relevance. Here, we describe a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identifying two human transforming growth factor beta1 (TGF-beta1) signal (leader) sequence polymorphisms, T869C (Leu10Pro) and G915C (Arg25Pro). This permits simple and robust identification of TGF-beta1 leader sequence genotypes and demonstrates the physical linkage in cis between T869C (Leu10Pro) and G915C (Arg25Pro). The method does not require previously genotyped standards. The efficacy of enzyme digestion is internally controlled by the presence of conserved restriction sites.
Wood
N A
NA
Department of Pathology and Microbiology, Homoeopathic Hospital Site, University of Bristol, Cotham, Bristol, UK.
Thomson
S C
SC
Smith
R M
RM
Bidwell
J L
JL
D001483:Base Sequence; D004247:DNA; D004262:DNA Restriction Enzymes / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D021382:Protein Sorting Signals / Q000235:genetics*; D016212:Transforming Growth Factor beta / Q000235:genetics*
D021382:Protein Sorting Signals; D016212:Transforming Growth Factor beta; D004247:DNA; D004262:DNA Restriction Enzymes