title: Expression, crystallization and preliminary X-ray crystallographic analysis of DNA-directed RNA polymerase subunit L from Thermococcus onnurineus NA1.
abstract: RNA polymerase (RNAP) plays a crucial role in gene expression in all organisms. It is a multiprotein complex that produces primary transcript RNA. Generally, the basal transcription apparatus in archaea is simpler than the eukaryotic RNA polymerase II counterpart. To understand the structure and function of archaeal RNAP, the TON-0309 gene encoding DNA-directed RNA polymerase subunit L (ToRNAP_L) from Thermococcus onnurineus NA1 was cloned and the protein was overexpressed in Escherichia coli, purified and crystallized. The purified protein was crystallized using the hanging-drop vapour-diffusion method and the crystal diffracted to 2.10 Å resolution. The crystal belonged to the hexagonal space group P6122, with unit-cell parameters a = b = 42.3, c = 211.2 Å. One molecule was present in the asymmetric unit, with a corresponding VM of 2.5 Å(3) Da(-1) and a solvent content of 50.0%.
authors.affiliation: Department of Biological Sciences, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Thien Hoang
authors.initials: TH
authors.lastname: Ho
authors.affiliation: Protein and Chemical Inc., Korea Industry and Academy Cooperation Building Room 814, 120 Neungdong-ro, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Myoung Ki
authors.initials: MK
authors.lastname: Hong
authors.affiliation: Department of Biological Sciences, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Ho Phuong Thuy
authors.initials: HP
authors.lastname: Ngo
authors.affiliation: Department of Biological Sciences, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Lin Woo
authors.initials: LW
authors.lastname: Kang
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D015971:Gene Expression Regulation, Enzymologic; D019712:Thermococcus / Q000201:enzymology*
chemical_list: D004247:DNA; D012321:DNA-Directed RNA Polymerases
keywords: DNA-directed RNA polymerase subunit L; Thermococcus onnurineus NA1