Alternative method for diagnosis of two polymorphisms in the human transforming growth factor-beta1 by PCR-mediated double site-directed mutagenesis.

Cytokine transforming growth factor-beta1 plays an important role in physiological processes during ontogenesis, cell differentiation, immune responses, carcinogenesis, inflammation, wound healing, fibroproduction, progression of renal insufficiency and arteriosclerotic lesion development. Its biological function is influenced through the two signal peptide polymorphisms. We describe a new, economical, easy and fast alternative method which allows detection of both polymorphisms from one PCR product with subsequent restriction analysis with two different restriction enzymes. This method could facilitate further research on the role of this cytokine in human disease.