1H38

Structure of a T7 RNA polymerase elongation complex at 2.9A resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SPRING-8 BEAMLINE BL45XU
Synchrotron site	SPring-8
Beamline	BL45XU
Temperature [K]	100
Detector technology	IMAGE PLATE
Detector	RIGAKU IMAGE PLATE
Spacegroup name	P 1
Unit cell lengths	79.907, 84.971, 202.003
Unit cell angles	90.36, 92.97, 109.94

Refinement procedure

Resolution	40.000 * - 2.900
R-factor	0.236
Rwork	0.236
R-free	0.28400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1qln
RMSD bond length	0.016
RMSD bond angle	21.500 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	2.690
High resolution limit [Å]	2.900 *	2.900 *
Rmerge	0.076 *	0.335 *
Total number of observations	348602 *
Number of reflections	108303 *
<I/σ(I)>	14.7	2.9
Completeness [%]	98.1 *	94.5 *
Redundancy	3.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, sitting drop *	8.1	20 *	10% PEG 8000, 8% GLYCEROL, 5MM BETA-MERCAPTOETHANOL,100 MM TRIS BUFFER, PH8.1, pH 8.10

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	10 (mg/ml)
2	1	reservoir	PEG8000	10 (%)
3	1	reservoir	glycerol	10 (%)
4	1	reservoir	beta-mercaptoethanol	5 (mM)
5	1	reservoir	Tris	100 (mM)	pH8.1