engleski

RFLP and SSCP of the HGF gene (IV exon PCR product)

Hepatocyte growth factor (HGF) is a multifunctional cytokine and a major factor of liver regeneration, regulating the cell growth, development, motility and morphogenesis. Active HGF consists of a (64 kDa) and b (34 kDa) chains linked by disulfide bond. The gene is located on chromosome 7, locus q 11.1-21, and consists of 18 exons and 17 introns of approximately 70 kb. The structure of HGF contains four kringle domains, each of them encoded by two exons. The first and second kringle domain, and hairpin loop are essential for HGF recognition by c-met/HGF receptors, as well as for HGF structural domain. The forth and fifth exons encode for the first structural domain. As mutations in functionally and biologically relevant domains lead to a pathologic process, polymorphism studies in healthy subjects and patients with liver diseases, and determination of their association with the occurrence of disease are of utmost importance. The polymorphism of DNA fragment obtained by polymerase chain reaction (PCR) can be determined by two established methods, i.e. restriction fragment length polymorphism (RFLP) and single-strand conformation polymorphism (SSCP). Here we described the PCR-RFLP and PCR-SSCP protocol for the polymorphism analysis of exon 4th of the HGF gene.

hepatocyte growth factor; HGF; polymorphysm; PCR-RFLP; PCR-SSCP exon 4th

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