Whole genome shotgun seqencing of Parageobacillus thermoglucosidasius NBRC107763 genetically engineered in carbon monoxide-dependent hydrogen production machinery

We deleted genes for carbon monoxide dehydrogenase, energy-converting hydrogenase, and both of genes in Parageobacillus thermoglucosidasius NBRC 107763 (DSM 2542). We then performed whole genome shotgun sequencing of wild type and three gene knockouts, and mapped the sequenced reads onto DSM 2542 complete genome sequence (NCBI RefSeq genome, NZ_CP012712) to support the gene deletions at the appropriate sites, to confirm no marker insertion into the genomes, and to identify unexpected gene mutations.

我们对热葡糖苷副芽孢杆菌（Parageobacillus thermoglucosidasius）NBRC 107763（DSM 2542）中编码一氧化碳脱氢酶（carbon monoxide dehydrogenase）、能量转换氢化酶（energy-converting hydrogenase）的基因，以及上述两类基因的双敲除组合进行了基因敲除。随后，我们针对野生型菌株与上述三株基因敲除菌株开展了全基因组鸟枪法测序（whole genome shotgun sequencing），并将所得测序读段（sequenced reads）比对至DSM 2542的完整基因组序列（NCBI参考序列基因组（NCBI RefSeq genome），登录号NZ_CP012712），以此验证靶位点基因敲除的正确性，确认基因组中未发生标记插入，同时识别出意外出现的基因突变。