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The standard procedure and reagents from the Q5 Site-Directed Mutagenesis kit (NEB) were used to insert mutations, ligate plasmids and transform them into NEB 5 competent cells by heat shock

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From an economic perspective, although crystalline amino acids can be costlier, their inclusion improves feed conversion ratio (FCR), enhances stress tolerance, and reduces mortality, yielding better productivity and resource efficiency [12], 105]