in vitro mutagenesis

Gene changes can help identify which genes are performing which functions.

process

Plasmids and other vectors can have genes. Added chemicals or enzymes can mutate genes {in vitro mutagenesis}. Vectors go into hosts, express genes, and make protein.

methods

Gene changes can be at restriction endonuclease sites. If sites have overhanging strands, S1 nuclease can remove overhanging single-strand DNA, or DNA polymerase can extend shorter strands, to make blunt ends. Linkers can attach to blunt ends.

Chemicals can alter gene nucleotides. Sodium bisulfite makes C into U. Hydrazine and formic acid delete nucleotide nitrogenous base, leaving sugar and phosphate. At low nucleotide concentrations or in harsh chemical conditions, DNA polymerase can add wrong nucleotides during DNA synthesis.

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