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However, we still can not jump to the conclusion that the strand-specific mutation pressure is the main force driving the evolution of tRNA anticodon for marine bivalve mitogenomes.

The mutation hypothesis of anticodon evolution argues that the strand-specific mutation pressure is the dominant force in shaping anticodon evolution and the anticodon wobble nucleotide bias should be in accord with the strand mutation bias.

Perhaps, in bacteria with larger chromosome, the mutation pressure is hard to prevail translational selection.

The asymmetrical directional mutation pressure is also reflected in the codon usage of genes oriented in opposite directions [ 48].

There is a significant positive correlation between GC12 and GC3 (r=0.394, p < 0.01), suggesting that the effect of directional mutation pressure is present at all codon positions.

This mutation pressure is applicable to CpG-containing codons because exons are generally methylated (Tornaletti and Pfeifer 1995; Rabinowicz et al. 2003).

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On the other hand, different selective mutation pressure was reported for genes located on different strands, and the genes on the lagging strand accumulate more mutations [11], [14].

Overall, mutation pressure was found to be the most important and dominant force that shapes codon usage patterns in MARV.

For instance, in the literature of molecular evolution, mutational explanations for patterns (e.g., GC content) are nearly universally described in terms of "mutation pressure", are labeled "neutral", and are considered rejected when there is evidence for selective allele fixations (e.g., [ 52, 53]).

The strength of the selection on a given gene relative to the mutation pressure were estimated by the method of the relative neutrality plot (RNP), which consists of plotting the G+C content at the nonsynonymous positions (GC12) of the codons against the G+C content at the synonymous position (GC3s).

Fourth, the scattering of the data points, which may reflect the interplay of selective constraints and mutation pressures, is rather impressive as some of the codons are almost invariable but others tend to go wild.

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