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Description And Function Of Mutagenesis

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Site-directed mutagenesis is a method used to construct amino acid changes in a protein in order to test the function of a specific amino acid. It is the way to study protein structure and function change the amino acid sequence of a protein by altering the DNA sequence of cloned gene4. A particular amino acid is very important in catalytic activity, ligand binding, protein folding or other function. Amino acid residue significance is tested by making conservative substitutions or by changing the amino acid to either alanine or glycine. Site-directed mutagenesis is also used to construct compensatory mutations, which are used to show the importance of specific interactions by making changes in both interacting partners. Each change individual destroy the interaction and combination of changes restores the interaction. These type of experiments are typically between two protein or between protein and nucleic acid. Used site-directed mutagenesis to investigate the interaction between hPol k and DNA1.

DNA polymerase kappa is a Y-family polymerase that use to bypass the damage DNA as translesion synthesis polymerase. In prostate cancer has reported that it has higher frequency of mutation in DNA polymerase kappa. 28% of patients have somatic mutations in POLK gene of prostate tumor and dominated by C- to –T transition2.

Approximately 30,000 DNA lesions occur per cell per day in mammalian cells. This damage to DNA can lead to cell death as well as disease such as cancer.

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