Identification of human TGF-beta 1 signal (leader) sequence polymorphisms by PCR-RFLP

Links between disease susceptibility and genetically determined variation i n human cytokine expression have recently been described. This has led to a demand for simple methods of identifying cytokine gene polymorphisms of po tential clinical relevance. Here, we describe a polymerase chain reaction-r estriction fragment length polymorphism (PCR-RFLP) method for identifying t wo human transforming growth factor beta 1 (TGF-beta 1) signal (leader) seq uence polymorphisms, T869C (Leu10Pro) and G915C (Arg25Pro). This permits si mple 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. Th e efficacy of enzyme digestion is internally controlled by the presence of conserved restriction sites. (C) 2000 Elsevier Science B.V. All rights rese rved.