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Saturation mutagenesis is commonly achieved by site-directed mutagenesis PCR with a randomised codon in the primers (e.g. SeSaM) [2] or by artificial gene synthesis, with a mixture of synthesis nucleotides used at the codons to be randomised. [3] Different degenerate codons can be used to encode sets of amino acids. [1]
Degeneracy or redundancy [1] of codons is the redundancy of the genetic code, exhibited as the multiplicity of three-base pair codon combinations that specify an amino acid. The degeneracy of the genetic code is what accounts for the existence of synonymous mutations . [ 2 ] :
Codon usage bias in Physcomitrella patens. Codon usage bias refers to differences in the frequency of occurrence of synonymous codons in coding DNA. A codon is a series of three nucleotides (a triplet) that encodes a specific amino acid residue in a polypeptide chain or for the termination of translation (stop codons).
With some exceptions, [1] a three-nucleotide codon in a nucleic acid sequence specifies a single amino acid. The vast majority of genes are encoded with a single scheme (see the RNA codon table ). That scheme is often called the canonical or standard genetic code, or simply the genetic code, though variant codes (such as in mitochondria ) exist.
The experiments used mixtures with all 20 amino acids. For each individual experiment, 19 amino acids were "cold" (nonradioactive), and one was "hot" (radioactively tagged with 14 C so they could detect the tagged amino acid later). They varied the "hot" amino acid in each round of the experiment, seeking to determine which amino acids would be ...
Early attempts at mutagenesis using radiation or chemical mutagens were non-site-specific, generating random mutations. [2] Analogs of nucleotides and other chemicals were later used to generate localized point mutations, [3] examples of such chemicals are aminopurine, [4] nitrosoguanidine, [5] and bisulfite. [6]
However, in a protein-coding sequence, the DNA sequence is made up of a string of codons which correspond to amino acids. Because the genetic code is degenerate (more than one codon map to a single amino acid) and samples from the amino acids rather than the codons, the codons are not sampled uniformly thus leading to differences in the PCFs.
Effective number of codons (abbreviated as ENC or Nc) is a measure to study the state of codon usage biases in genes and genomes. [1] The way that ENC is computed has obvious similarities to the computation of effective population size in population genetics. [2]