Identification of human TGF-β1 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 beta 1 (TGF-beta 1) signal (leader) sequence polymorphisms, T869C (Leu10Pro) and G915C (Arg25Pro). This permits simple and robust identification of TGF-beta 1 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. (C) 2000 Elsevier Science B.V. All rights reserved.