pBJG1 T7-8xHis-OGT D554N
(Plasmid
#154279)
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PurposeBacterial expression of full length human OGT with D554N mutation (Cleavage, no Ser/Thr glycosylation). N-terminal T7_leader-8xHis tag followed by HRV3C protease site.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 154279 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET24b(+)
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Backbone manufacturerNovagen
- Backbone size w/o insert (bp) 5000
- Total vector size (bp) 8412
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Modifications to backbonesubcloned MCS from alternate pET vector to take tag as described in SI of Gross et al., JACS 2005
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)NovaBlue
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameO-GlcNAc Transferase
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Alt nameOGT
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SpeciesH. sapiens (human)
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Insert Size (bp)3135
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MutationAspartate 554 changed to Asparagine- bacterial codon optimized
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Entrez GeneOGT (a.k.a. HINCUT-1, HRNT1, MRX106, O-GLCNAC, OGT1, XLID106)
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Tags
/ Fusion Proteins
- T7 leader (N terminal on insert)
- 8x His (N terminal on insert)
- HRV3C protease site (N terminal on insert)
Resource Information
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Supplemental Documents
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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.
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For your Materials & Methods section:
pBJG1 T7-8xHis-OGT D554N was a gift from Suzanne Walker (Addgene plasmid # 154279 ; http://n2t.net/addgene:154279 ; RRID:Addgene_154279) -
For your References section:
How the glycosyltransferase OGT catalyzes amide bond cleavage. Janetzko J, Trauger SA, Lazarus MB, Walker S. Nat Chem Biol. 2016 Nov;12(11):899-901. doi: 10.1038/nchembio.2173. Epub 2016 Sep 12. 10.1038/nchembio.2173 PubMed 27618188