Expression analysis of Thermococcus onnurineus NA1 KCTC10859 and an Frh-deficient mutant, Frh overexpression mutant and TON_0282 deletion mutant in a T. onnurineus NA1 strain.

Investigation of whole genome gene expression level changes in an Frh-deficient mutant, Frh overexpression mutant and TON_0282 deletion mutant, compared to the wild-type strain. In this study, as a first step to assess the role of the frhAGB-encoding hydrogenases and TON_0282 gene, we constructed three mutants by deleting each gene in a T. onnurineus NA1. A custom microarray was manufactured by Roche NimbleGen. Unique six 60-mer oligonucleotides for all of the predicted 1,986 open reading frames (ORFs) in the annotated genome of T. onnurineus NA1 were designed and synthesized. Examined Wildtype, Frh-deficient mutant, Frh overexpression mutant and TON_0282 deletion mutant in T. onnurineus NA1 strain.

本研究以野生型菌株为对照，探究了Frh缺陷突变株、Frh过表达突变株以及TON_0282缺失突变株的全基因组基因表达水平变化。为评估编码frhAGB的氢化酶与TON_0282基因的功能，本研究作为相关前期工作的第一步，在onnurineus NA1菌株（T. onnurineus NA1）中分别敲除各目标基因，成功构建三株突变体。罗氏尼姆布基因（Roche NimbleGen）定制制备了基因芯片（custom microarray），针对该菌株注释完成的基因组中预测得到的1986个开放阅读框（open reading frames, ORFs），为每一个开放阅读框设计并合成了6条独特的60聚体寡核苷酸探针。本研究对该菌株的野生型菌株、Frh缺陷突变株、Frh过表达突变株以及TON_0282缺失突变株进行了检测分析。