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Present challenges of
experimenting with plasmid systems

1. 1.

Plasmid replication

Origin of replication (Ori) is where the plasmid replication is initiated. Plasmids with same Oris use the same replication machinery.

Problem #1

1. 2.

Unequal replication

Plasmids with growth advantages such as faster replication (due to their smaller size) have the potential to rapidly outgrow larger plasmids in the cell.

Problem #1

1. 3.

Plasmid loss

This is even more likely to occur when the plasmid copy number is low. When the cell divides, lower copy plasmids tend to be lost in next generations.

Problem #1

2. 1.

Plasmid incompatibility

One needs to use different types of Ori to avoid this trouble, however it often causes dilemma due to limited selection options or the lack of good combination of Ori and plasmid‘s copy number.

Problem #2

2. 2.

Pavadinimas

Imagine you require a system consisting of two - low and high copy plasmids. Try picking them from the list above and see what happens.

Problem #2

2. 3.

Pavadinimas

Currently no handy method of changing the copy number of particular plasmid exists.

Problem #2

2. 4.

Pavadinimas

There is also no mechanism to adjust the copy number when so is required in any specific situation: for example, incoming cellular signals or special patterns.