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THEME 5 GENETICS

Methods: Screening of the ANG gene: We scanned the coding regions of the ANG gene by high resolution melting (HRM) analysis and sequenced the samples indicated to include mutations. Synthesis of ANG protein: We constructed mammalian expression plasmid vectors with C terminal FLAG tags including wild and mutated type ANG genes. The resultant plasmid DNAs were transfected into HeLa cells, and then angiogenin protein synthesis was examined by Western blotting and immunocytochemistry.