Revision as of 13:12, 1 November 2017

Rop protein - SynORI Inducible global copy number regulator(All groups)

This device is a fully functional inducible global copy number regulator that works with SynORI framework and other systems based on ColE1 origin of replication. It consists of Rhamnose promoter and ROP protein.

With this device plasmid copy number can be lowered globally (device affects every plasmid group of SynORI system) by increasing rhamnose concentration and therefor increasing ROP protein expression

See how this part fits into the whole SynORI framework by pressing here!

Sequence and Features

Figure 1. Main principles of ColE1 plasmid family replication. Rop protein interaction region marked in red square. (Citation needed)

Introduction

Biology

Repressor of primer (ROP) is a small dimeric protein that participates in ColE1 plasmid family copy number control, by increasing affinity between two complementary RNAs - RNA I (Replication inhibitor) and RNA II (Replication activator) (Fig. 1). ^[1] By increasing affinity of the two RNA molecules, Rop decreases the rate of plasmid replication initiation events.

Figure 2. Structure of the ColE1 Rop protein, at 1.7 angstroms resolution.^[2]

Rop dimer is a bundle of four tightly packed alpha helices that are held by hydrophobic interactions (Fig. 2).

Usage with SynORI (Framework for multi-plasmid systems)

About SynORI

SynORI is a framework for multi-plasmid systems created by Vilnius-Lithuania 2017 which enables quick and easy workflow with multiple plasmids, while also allowing to freely pick and modulate copy number for every unique plasmid group! Read more about [http://2017.igem.org/Team:Vilnius-Lithuania SynORI here]!

This device in SynORI

This is a inducible copy number device which sets a specific copy number for each plasmid group irrespective of groups present in the system. ROP protein expression is induced by different concentrations of rhamnose, therefor allowing to change the plasmid copy number when required without the need to build new system.

Further details

For more background information and indepth insight on this part's design please see the individual part page of ROP protein part:BBa_K2259010.

See the [http://2017.igem.org/Team:Vilnius-Lithuania Vilnius-Lithuania 2017 team wiki] for more insight about our synthetic origin of replication (SynORI).

Characterization of RNA II (Vilnius-Lithuania 2017)

Plasmid with SynORI constitutive number devices copy calculation

References

1. ↑ Castagnoli L, Scarpa M, Kokkinidis M, Banner DW, Tsernoglou D, Cesareni G. Genetic and structural analysis of the ColE1 Rop (Rom) protein. The EMBO Journal. 1989;8(2):621-629.
2. ↑ Banner DW, Kokkinidis M, Tsernoglou D. Structure of the ColE1 Rop protein at 1.7 Å resolution. J Mol Biol. 1987 m.;196(3):657–75.