Line 48: | Line 48: | ||
<div class="step">2. 3.</div> | <div class="step">2. 3.</div> | ||
<div class="title">Fixed Copy Number</div> | <div class="title">Fixed Copy Number</div> | ||
− | <p>Currently no handy method of changing the copy number of particular plasmid exists. It | + | <p>Currently no handy method of changing the copy number of particular plasmid exists. It must be either low, or medium, or high.</p> |
<h6>Problem #2</h6> | <h6>Problem #2</h6> | ||
</div> | </div> |
Latest revision as of 03:01, 1 November 2017
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Present Challenges of
Experimenting With Plasmid Systems
Origin of replication (Ori) is a sequence where the plasmid replication begins. Plasmids with similar Ori sites use the same replication machinery.
Problem #1
Replication machinery does not discriminate between similar Ori sites, therefore smaller plasmids are replicated faster and rapidly outgrow larger plasmids, which leads to uneven plasmid ratios in a cell.
Problem #1
This is even more likely to occur when some plasmids are low copy number. When the cell divides, low copy plasmids tend to be lost in next generations.
Problem #1
This inability to stably co-maintain different plasmids over a number of generations in the same bacterial cell line is called plasmid incompatibility. One needs to use different types of Ori to avoid this trouble.
Problem #2
However, it often causes trouble due to limited Ori selection options or the lack of good combination of Ori and plasmid's copy number. Imagine you require a system consisting of two - low and high copy number plasmids. Try picking them from the list.
Problem #2
Currently no handy method of changing the copy number of particular plasmid exists. It must be either low, or medium, or high.
Problem #2
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.
Problem #2
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A
![](https://static.igem.org/mediawiki/2017/3/39/T--Vilnius-Lithuania--step-2-1-plasmid1.png)
B
![](https://static.igem.org/mediawiki/2017/b/b4/T--Vilnius-Lithuania--step-2-1-plasmid2.png)
C
![](https://static.igem.org/mediawiki/2017/1/1b/T--Vilnius-Lithuania--step-1-2-boxline2.png)
D
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Plasmid
Copy number
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