|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||32102||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerNew England Biolabs
- Backbone size w/o insert (bp) 6700
Modifications to backboneAlready has one copy of MT inserted to begin with. This parent plasmid is available as Addgene plasmid 32101.
Vector typeBacterial Expression
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameMT (2x)
SpeciesM. musculus (mouse)
Entrez GeneMt1 (a.k.a. MT-I, Mt-1)
- Promoter P-lac
/ Fusion Protein
- MBP (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site XmnI (not destroyed)
- 3′ cloning site AflIII/NcoI (destroyed during cloning)
- 5′ sequencing primer MBP_F (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byThe metallothionein (MT) gene was amplified from pET-3d-MT (a gift from the Winge laboratory, University of Utah), which contains the MT gene within the pET-3d vector (Novagen, Madison, WI).
Terms and Licenses
- Not Available to Industry
Article Citing this Plasmid
In this construct, a translational modification was made that changes an alanine to an aspartic acid at the junction between the two MT genes. The second copy of the MT gene was inserted into the pMAL-c2x-MT1 plasmid (Addgene plasmids 32101) before the existing MT copy. The inserted MT gene was amplified from pET-3d-MT using the forward primer (50-CTCGGGATCGAGGGAAGGATTTCAAGATATACCATGGACCCC-30), which codes for the MBP linker region containing an XmnI and NcoI site fused in frame to the MT gene, and the reverse primer (50-GTGACCACATGTCACAGCACGTGCACTTGTCC-3 0 ), which contains an AflIII site at the MT–MT junction. The pMAL-c2x-MT plasmid was prepared by digesting with XmnI and NcoI, and the PCR product was constructed with XmnI and AflIII. Since NcoI and AflIII produced equivalent DNA ends, the second copy of MT was inserted with removal of both restriction sites at the ligation junction. This process can be reproduced iteratively to introduce as many copies of MT in frame into the construct as desired. In addition to this plasmid Addgene offers versions with 1 copy (32101), 3 copies (32115) and 4 copies (32100) of MT.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pMAL-c2x-MT2 was a gift from David DeRosier & Chris Mercogliano & Cristina Risco Ortiz (Addgene plasmid # 32102 ; http://n2t.net/addgene:32102 ; RRID:Addgene_32102)
For your References section:Concatenated metallothionein as a clonable gold label for electron microscopy. Mercogliano CP, DeRosier DJ. J Struct Biol. 2007 Oct;160(1):70-82. Epub 2007 Jul 10. 10.1016/j.jsb.2007.06.010 PubMed 17692533