Difference between revisions of "Team:HokkaidoU Japan/his-tag"
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<h2 id="overview">Overview</h2> | <h2 id="overview">Overview</h2> | ||
| − | + | <p> One of the ways to produce large quantities of a recombinant toxic protein in E.coli is to aggregate it as inclusion bodies (IBs). IBs are insoluble protein aggregations observed in bacteria overexpressing heterologous genes whose products fail to attain a soluble (de Groot et al. 2008). Protein aggregation can lose the activity of the protein to some extent. So we optimized inclusion bodies to achieve large amounts of the toxic peptide, thanatin.<br></p> | |
| + | <p> It’s known that using a histidine tag as a fusion partner for protein expression did affect the formation of inclusion bodies. In our project, we intended to express thanatin with a 10x His-tag at the N terminus as an inclusion body, then cleave of a His-tag with some proteinase (TEV proteinase) leading successfully to express thanatin in a soluble form.</br> | ||
| + | <ul> | ||
| + | <li>Inclusion bodies (IBs)</li> | ||
| + | <li> His tag</li> | ||
| + | <li>TEV poteinase</li> | ||
| + | </ul></p> | ||
<h2 id="design">Design</h2> | <h2 id="design">Design</h2> | ||
Revision as of 01:19, 15 September 2015
His-tag Inactivation System
Overview
One of the ways to produce large quantities of a recombinant toxic protein in E.coli is to aggregate it as inclusion bodies (IBs). IBs are insoluble protein aggregations observed in bacteria overexpressing heterologous genes whose products fail to attain a soluble (de Groot et al. 2008). Protein aggregation can lose the activity of the protein to some extent. So we optimized inclusion bodies to achieve large amounts of the toxic peptide, thanatin.
It’s known that using a histidine tag as a fusion partner for protein expression did affect the formation of inclusion bodies. In our project, we intended to express thanatin with a 10x His-tag at the N terminus as an inclusion body, then cleave of a His-tag with some proteinase (TEV proteinase) leading successfully to express thanatin in a soluble form.
- Inclusion bodies (IBs)
- His tag
- TEV poteinase
Design
Experiments
Result
Conclusion
