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.. Thermococcus onnurineus NA1

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. Overall design: 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.

本研究针对Frh缺陷突变株、Frh过表达突变株及TON_0282基因缺失突变株与野生型菌株的全基因组基因表达水平变化展开分析。为评估编码frhAGB的氢化酶以及TON_0282基因的功能，本研究作为前期探索步骤，在T. onnurineus NA1菌株中分别敲除对应基因，构建了上述三株突变体。
整体实验设计：由罗氏NimbleGen（Roche NimbleGen）定制制备基因芯片。针对T. onnurineus NA1已注释基因组中预测的1986个开放阅读框（open reading frame, ORF），为每个开放阅读框设计并合成6条独特的60聚体寡核苷酸探针。本实验检测的样本包括T. onnurineus NA1的野生型菌株、Frh缺陷突变株、Frh过表达突变株及TON_0282缺失突变株。