Gene Cloning and Function Analysis of Replication Factor C fromThermococcus kodakaraensisKOD1 | Zendy

Masao Kitabayashi | Zendy; Yoshiaki Nishiya | Zendy; Muneharu Esaka | Zendy; Mitsuo Itakura | Zendy; Tadayuki Imanaka | Zendy

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Gene Cloning and Function Analysis of Replication Factor C fromThermococcus kodakaraensisKOD1

Author(s) -

Masao Kitabayashi,

Yoshiaki Nishiya,

Muneharu Esaka,

Mitsuo Itakura,

Tadayuki Imanaka

Publication year - 2003

Publication title -

bioscience biotechnology and biochemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.509

H-Index - 116

eISSN - 1347-6947

pISSN - 0916-8451

DOI - 10.1271/bbb.67.2373

Subject(s) - thermococcus , biology , dna polymerase , dna polymerase delta , dna clamp , microbiology and biotechnology , dna replication , polymerase , dna polymerase ii , proliferating cell nuclear antigen , dna , gene , genetics , polymerase chain reaction , reverse transcriptase , archaea

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.

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