Difference between revisions of "Plasmid backbones/Assembly of protein fusions"

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Protein engineers often need to do in-frame assemblies in order to assemble together different signal sequences, proteins domains, and protein tags.  BioBrick&trade; standard assembly is not well-designed for this task because the scar sequence formed by the SpeI-XbaI ligation is 8 base pairs long.  As a result, different groups have developed modified or alternative assembly schemes to facilitate assembly of protein domains.
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Pam Silver's lab has developed the Silver lab standard for assembling protein domains.  It relies on shortening the BioBrick prefix and suffix each by 1 base pair such that the resulting SpeI-XbaI scar is only 6 base pairs long and protein domains can be assembled in frame.
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<parttable>assembly_fusion_plasmid</parttable>
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#assembly_fusion_plasmid_table th {background-color: #aaaaaa;padding: 5px; font-size:14px;}
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===References===
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'''A New Biobrick Assembly Strategy Designed for Facile Protein Engineering'''<br>
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''MIT SBWG Technical Reports,'' 2006 Apr 20<br>
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Ira Philips, Pam Silver<br>
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[http://hdl.handle.net/1721.1/32535 URL] (open access!)

Revision as of 20:11, 1 August 2008

< Back to Plasmids

Protein engineers often need to do in-frame assemblies in order to assemble together different signal sequences, proteins domains, and protein tags. BioBrick™ standard assembly is not well-designed for this task because the scar sequence formed by the SpeI-XbaI ligation is 8 base pairs long. As a result, different groups have developed modified or alternative assembly schemes to facilitate assembly of protein domains.

Pam Silver's lab has developed the Silver lab standard for assembling protein domains. It relies on shortening the BioBrick prefix and suffix each by 1 base pair such that the resulting SpeI-XbaI scar is only 6 base pairs long and protein domains can be assembled in frame.


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References

A New Biobrick Assembly Strategy Designed for Facile Protein Engineering
MIT SBWG Technical Reports, 2006 Apr 20
Ira Philips, Pam Silver
[http://hdl.handle.net/1721.1/32535 URL] (open access!)