pGEX6p1_GST-HRV3C-DmrB-[dltCARD]mCasp1(E102D103TEV)-8xHis (KS585)
(Plasmid
#254943)
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PurposeMurine caspase-1 with the E102/D103 autocleavage site replaced by a TEV site, CARD replaced by DmrB, N-terminal GST followed by a PreScission (HRV3C) site, and a C-terminal 8xHis tag.
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Depositing Lab
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 254943 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
Backbone
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Vector backbonepGEX6p1
- Backbone size w/o insert (bp) 4966
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameCasp1
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Alt nameInterleukin-1 beta convertase (Il1bc)
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Alt nameInterleukin-1 beta-converting enzyme (ICE)
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SpeciesM. musculus (mouse)
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Insert Size (bp)1296
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MutationE102D103TEV
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Entrez GeneCasp1 (a.k.a. ICE, Il1bc)
- Promoter tac
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Tags
/ Fusion Proteins
- GST (N terminal on backbone)
- 8xHis (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site XhoI (not destroyed)
- 5′ sequencing primer pGEX_MCS_seq_fw (GGGCTGGCAAGCCACGTTTGGTG)
- 3′ sequencing primer pGEX_MCS_seq_rv (CACCGAAACGCGCGAGGCAGATC)
- (Common Sequencing Primers)
Resource Information
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.
Depositor Comments
Insert has a STOP but no START (N-terminal GST fusion).
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:
pGEX6p1_GST-HRV3C-DmrB-[dltCARD]mCasp1(E102D103TEV)-8xHis (KS585) was a gift from Kate Schroder (Addgene plasmid # 254943 ; http://n2t.net/addgene:254943 ; RRID:Addgene_254943) -
For your References section:
Caspase-1 self-cleavage is an intrinsic mechanism to terminate inflammasome activity. Boucher D, Monteleone M, Coll RC, Chen KW, Ross CM, Teo JL, Gomez GA, Holley CL, Bierschenk D, Stacey KJ, Yap AS, Bezbradica JS, Schroder K. J Exp Med. 2018 Mar 5;215(3):827-840. doi: 10.1084/jem.20172222. Epub 2018 Feb 6. 10.1084/jem.20172222 PubMed 29432122