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− | <h1>Present | + | <h1>Present Challenges of<br/> Experimenting With Plasmid Systems</h1> |
<div class="presentation"> | <div class="presentation"> | ||
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<div class="left-step is-current" style="display: block"> | <div class="left-step is-current" style="display: block"> | ||
<div class="step">1. 1.</div> | <div class="step">1. 1.</div> | ||
− | <div class="title">Plasmid | + | <div class="title">Plasmid Replication </div> |
<p>Origin of replication (Ori) is a sequence where the plasmid replication begins. Plasmids with similar Ori sites use the same replication machinery. </p> | <p>Origin of replication (Ori) is a sequence where the plasmid replication begins. Plasmids with similar Ori sites use the same replication machinery. </p> | ||
<h6>Problem #1</h6> | <h6>Problem #1</h6> | ||
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<div class="left-step"> | <div class="left-step"> | ||
<div class="step">1. 2.</div> | <div class="step">1. 2.</div> | ||
− | <div class="title"> | + | <div class="title">Unbalanced Replication</div> |
− | <p> Replication machinery does not discriminate between similar Ori sites, therefore smaller plasmids are replicated faster and rapidly | + | <p> 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.</p> |
<h6>Problem #1</h6> | <h6>Problem #1</h6> | ||
</div> | </div> | ||
<div class="left-step"> | <div class="left-step"> | ||
<div class="step">1. 3.</div> | <div class="step">1. 3.</div> | ||
− | <div class="title">Plasmid | + | <div class="title">Plasmid Loss</div> |
− | <p> This is even more likely to occur when | + | <p> 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. </p> | divides, low copy plasmids tend to be lost in next generations. </p> | ||
<h6>Problem #1</h6> | <h6>Problem #1</h6> | ||
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<div class="left-step"> | <div class="left-step"> | ||
<div class="step">2. 1.</div> | <div class="step">2. 1.</div> | ||
− | <div class="title">Plasmid | + | <div class="title">Plasmid Incompatibility </div> |
− | <p>One needs to use different types of Ori to avoid this trouble | + | <p>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.</p> |
− | + | ||
− | + | ||
<h6>Problem #2</h6> | <h6>Problem #2</h6> | ||
</div> | </div> | ||
<div class="left-step"> | <div class="left-step"> | ||
<div class="step">2. 2.</div> | <div class="step">2. 2.</div> | ||
− | <div class="title"> | + | <div class="title">Plasmid Incompatibility</div> |
− | <p>Imagine you require a system consisting of two - low and high copy plasmids. Try picking them | + | <p>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.</p> |
− | + | ||
<h6>Problem #2</h6> | <h6>Problem #2</h6> | ||
</div> | </div> | ||
<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. It must be either low, or medium, or high.</p> |
<h6>Problem #2</h6> | <h6>Problem #2</h6> | ||
</div> | </div> | ||
<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 | + | situation, for example: incoming cellular signals or special patterns.</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
Did this look smart, easy or convenient?
We think it is time for a change.
A
B
C
D
Plasmid
Copy number