Characterization of the emergent properties of a synthetic quasi-cellular system

BMC Bioinformatics

Table 1 PURE system components*

Component	Conc.	Component	Conc.
Translation factors		ProRS	1,300 U/mL
IF1	2.7 μM	SerRS	1,900 U/mL
IF2	0.4 μM	ThrRS	1,300 U/mL
IF3	1.5 μM	TrpRS	630 U/mL
EF-G	0.26 μM	TyrRS	630 U/mL
EF-Tu	0.92 μM	ValRS	3,100 U/mL
EF-Ts	0.66 μM
RF1	0.25 μM	Other enzymes
RF2	0.24 μM	MTF	4,500 U/mL
RF3	0.17 μM	Ribosomes	1.2 μM
RRF	0.5 μM	Creatine kinase	4 μg/mL
		Myokinase	3 μg/mL
Amino acyl-tRNA synthetases (RSs)		Nucleoside diphosphate kinase	1.1 μg/mL
AlaRS	1,900 U/mL	Pyrophosphatase	2 U/mL
ArgRS	2,500 U/mL	T7 RNA polymerase	10 μg/mL
AsnRS	20 mg/mL
AspRS	2,500 U/mL	Energy sources
CysRS	630 U/mL	ATP	2 mM
GlnRS	1,300 U/mL	GTP	2 mM
GluRS	1,900 U/mL	CTP	1 mM
GlyRS	5,000 U/mL	UTP	1 mM
HisRS	630 U/mL	Creatine phosphate	20 mM
IleRS	2,500 U/mL
LeuRS	3,800 U/mL	Other components
LysRS	3,800 U/mL	20 amino acids	0.3 mM
MetRS	6,300 U/mL	10-formyl-5,6,7,8-tetrahydrofolic acid	10 mg/mL
PheRS	1,300 U/mL	tRNAmix (Roche)	56 Abs260

*The components are solubilized in 50 mM HEPES-KOH pH 7.6; 100 mM potassium glutamate, 13 mM magnesium acetate, 2 mM spermidine, 1 mM DTT. One unit of activity was defined as the amount of enzyme that catalyzes the formation of 1 pmol of amino acyl-tRNA in 1 min (reproduced from [35] with permission from Elsevier). The conversion from U/mL to μM has been taken from [27].

ISSN: 1471-2105