PTXB1

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'''Interested in adding a page on a material to OpenWetWare?  Here is a template to help you do so with some suggested headings.  Please use or modify as you see fit.  Not all the headings will necessarily be relevant to your material.  Feel free to add or delete sections as appropriate.'''
 
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'''Click the view source tab and copy everything below this line.  Paste it into your new material page.  Then replace the text in this page with your own material's information.'''
 
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<!-- COPY EVERYHING BELOW HERE TO START YOUR OWN MATERIAL!  -->
 
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==Purpose==
==Purpose==
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[[''E. coli'']] [[cloning vector]] that creates a C-terminal fusion to an [[intein]] domain, which is useful in techniques such as [[expressed protein ligation]].
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''[[E. coli]]'' [[cloning vector]] that creates a C-terminal fusion to an [[intein]] domain, which is useful in techniques such as [[expressed protein ligation]].
==Procurement==
==Procurement==
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[[http://neb.com|New England Biolabs]]
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Sold by [http://neb.com| New England Biolabs].
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==Map, Sequence and MCS==
==Use==
==Use==
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*How does one use this material.
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Clone a gene using appropriate restriction sites into the [[Multiple Cloning Site|MCS]]. Protein expressed from this plasmid in ''[[E. coli]]'' can be purified on a chitin affinity column. The tag can be cleaved on-column with reducing agents such as [[dithiothreitol|DTT]] or [[MESNA]], leaving a C-terminal reactive thioester. Peptides and proteins with N-terminal [[cysteine|Cys]] residues will react with the thioester to form a native peptide bond.
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*Include any relevant links to [[Protocols|OWW protocols]]
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This vector is part of the IMPACT-CN kit:
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*Include any relevant links to the manufacturer's protocols
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* [http://www.neb.com/nebecomm/ManualFiles/manualE6901.pdf| IMPACT-CN manual]
==Safety==
==Safety==
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*Are there any safety considerations for this material?
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N/A
==References==
==References==
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*Any papers or websites that are relevant to the use of this material
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<PMID>pTXB1 or IMPACT</PMID>
==Contact==
==Contact==
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*Who has experience using this material?
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[[User:Kathryn Muratore|Kathryn Muratore]]
==Categories==
==Categories==
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Add any other categories that may be relevant to your page.  We use categories like tags.
 
[[Category:Material]]
[[Category:Material]]
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[[Category:Miscellaneous]]
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[[Category:Escherichia coli]]
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[[Category:Cloning Vector]]
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*Please replace the second category (Miscellaneous) with one or more category tags that identifies where in the Material hierarchy this page belongs. Remove the 'Miscellaneous' category to keep it from ending back up in the list of structural pages that make up that category.
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Revision as of 14:04, 25 May 2011

Contents

Purpose

E. coli cloning vector that creates a C-terminal fusion to an intein domain, which is useful in techniques such as expressed protein ligation.

Procurement

Sold by New England Biolabs.

Map, Sequence and MCS

Use

Clone a gene using appropriate restriction sites into the MCS. Protein expressed from this plasmid in E. coli can be purified on a chitin affinity column. The tag can be cleaved on-column with reducing agents such as DTT or MESNA, leaving a C-terminal reactive thioester. Peptides and proteins with N-terminal Cys residues will react with the thioester to form a native peptide bond. This vector is part of the IMPACT-CN kit:

Safety

N/A

References

<PMID>pTXB1 or IMPACT</PMID>

Contact

Kathryn Muratore

Categories

Personal tools