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Mycoplasma play important roles in the emerging field synthetic biology due to their small genomes and simple metabolic pathways.The distribution of homologous essential genes among different Mycoplasma species is crucial to the design of the ideal chassis.Therefore,a gene essentiality prediction algorithm has been developed,which incorporates information of biased gene strand distribution,homology search and codon adaptation index.Furthermore,this in-house bioinformatics algorithm has been used to predict 7,676 essential genes from 22 representative Mycoplasma genomes.