Kitabayashi Masao, Nishiya Yoshiaki, Esaka Muneharu, Itakura Mitsuo, Imanaka Tadayuki
Tsuruga Institute of Biotechnology, Toyobo Co., Ltd., Tsuruga, Fukui, Japan.
Biosci Biotechnol Biochem. 2003 Nov;67(11):2373-80. doi: 10.1271/bbb.67.2373.
Replication factor C (RFC) catalyzes the assembly of circular proliferating cell nuclear antigen (PCNA) clamps around primed DNA, enabling processive synthesis by DNA polymerase. The RFC-like genes, arranged in tandem in the Thermococcus kodakaraensis KOD1 genome, were cloned individually and co-expressed in Escherichia coli cells. T. kodakaraensis KOD1 RFC homologue (Tk-RFC) consists of the small subunit (Tk-RFCS: MW=37.2 kDa) and the large subunit (Tk-RFCL: MW=57.2 kDa). The DNA elongation rate of the family B DNA polymerase from T. kodakaraensis KOD1 (KOD DNA polymerase), which has the highest elongation rate in all thermostable DNA polymerases, was increased about 1.7 times, when T. kodakaraensis KOD1 PCNA (Tk-PCNA) and the Tk-RFC at the equal molar ratio of KOD DNA polymerase were reacted with primed DNA.
复制因子C（RFC）催化在引发的DNA周围组装环状增殖细胞核抗原（PCNA）夹子，从而使DNA聚合酶能够进行持续合成。在嗜热栖热菌KOD1基因组中串联排列的类RFC基因被单独克隆并在大肠杆菌细胞中共表达。嗜热栖热菌KOD1 RFC同源物（Tk-RFC）由小亚基（Tk-RFCS：分子量 = 37.2 kDa）和大亚基（Tk-RFCL：分子量 = 57.2 kDa）组成。当嗜热栖热菌KOD1 PCNA（Tk-PCNA）和与KOD DNA聚合酶等摩尔比的Tk-RFC与引发的DNA反应时，来自嗜热栖热菌KOD1的B族DNA聚合酶（KOD DNA聚合酶）的DNA延伸率在所有耐热DNA聚合酶中具有最高的延伸率，提高了约1.7倍。
