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<div class="left-step"> | <div class="left-step"> | ||
<div class="step">2. 3.</div> | <div class="step">2. 3.</div> | ||
− | <div class="title"> | + | <div class="title">Fixed copy number</div> |
<p>Currently no handy method of changing the copy number of particular plasmid exists.</p> | <p>Currently no handy method of changing the copy number of particular plasmid exists.</p> | ||
<h6>Problem #2</h6> | <h6>Problem #2</h6> | ||
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<div class="left-step"> | <div class="left-step"> | ||
<div class="step">2. 4.</div> | <div class="step">2. 4.</div> | ||
− | <div class="title"> | + | <div class="title">Deficient responsivity</div> |
<p>There is also no mechanism to adjust the copy number when so is required in any specific | <p>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.</p> | situation: for example, incoming cellular signals or special patterns.</p> |
Revision as of 22:16, 31 October 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 one plasmid is 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 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.
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
Did this look smart, easy or convenient?
We think it is time for a change.
A
B
C
D
Plasmid
Copy number