PTXB1: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(New page: <center> {|style="width: 48em; background: #FF6677;" | align="center"|'''This page is a template and should not be edited.'''<br><span style="font-size:90%">Click [http://openwetware.org/i...) |
|||
| (6 intermediate revisions by the same user not shown) | |||
| Line 1: | Line 1: | ||
==Purpose== | ==Purpose== | ||
[[ | ''[[E. coli]]'' cloning [[vectors|vector]] that creates a C-terminal fusion to an [[intein]] domain, which is useful in techniques such as [[expressed protein ligation]]. | ||
==Procurement== | ==Procurement== | ||
Sold by [http://neb.com| New England Biolabs]. | |||
==Map, Sequence and MCS== | |||
[[Image:PTXB1.svg|600px]] | |||
*[http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/sequences/Fasta/pTXB1.fsa.txt| Sequence as FASTA] | |||
*[http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/sequences/GenBank/pTXB1.gbk.txt| Sequence in genbank format] | |||
*[http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/maps/pTXB1_map.pdf| Technical reference including MCS, map, and other information] | |||
==Use== | ==Use== | ||
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. | |||
This vector is part of the IMPACT-CN kit: | |||
* | * [http://www.neb.com/nebecomm/ManualFiles/manualE6901.pdf| IMPACT-CN manual] | ||
==Safety== | ==Safety== | ||
N/A | |||
==References== | ==References== | ||
==Contact== | ==Contact== | ||
[[User:Kathryn Muratore|Kathryn Muratore]] | |||
==Categories== | ==Categories== | ||
[[Category:Material]] | [[Category:Material]] | ||
[[Category: | [[Category:Escherichia coli]] | ||
[[Category:DNA]] | |||
[[Category:Cloning Vector]] | |||
Latest revision as of 15:07, 25 May 2011
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
- Sequence as FASTA
- Sequence in genbank format
- Technical reference including MCS, map, and other information
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
