Identification of Essential Genes in Streptococcus pneumoniae by Allelic Replacement Mutagenesis

作者：Jae-Hoon Song, Kwan Soo Ko, Jiyoung Lee, Jin Yang Baek, Won Sup Oh, Ha Sik Yoon, Jin‐Yong Jeong, Jongsik Chun · 发表于：Molecules and Cells · 年份：2005 · DOI：10.1016/s1016-8478(23)13181-5 · 被引用次数：200 · 研究领域：Genomics and Phylogenetic Studies、Pneumonia and Respiratory Infections、RNA and protein synthesis mechanisms

To find potential targets of novel antimicrobial agents, we identified essential genes of Streptococcus pneumoniae using comparative genomics and allelic replacement mutagenesis. We compared the genome of S. pneumoniae R6 with those of Bacillus subtilis, Enterococcus faecalis, Escherichia coli, and Staphylococcus aureus, and selected 693 candidate target genes with > 40% amino acid sequence identity to the corresponding genes in at least two of the other species. The 693 genes were disrupted and 133 were found to be essential for growth. Of these, 32 encoded proteins of unknown function, and we were able to identify orthologues of 22 of these genes by genomic comparisons. The experimental method used in this study is easy to perform, rapid and efficient for identifying essential genes of bacterial pathogens.