Cer(1-158)-gamma-1 in pcDNAI/Amp
(Plasmid
#55623)
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PurposeAn amino-terminal mCerulean fragment was fused to Ggamma-1. When co-expressed with a carboxyl-terminal CFP fragment fused to a Gbeta subunit with which it interacts, a fluorescent signal is produced.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 55623 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNAI/Amp
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 4800
- Total vector size (bp) 5476
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Modifications to backboneA BglII site was introduced into the polylinker of pcDNAI/Amp 3' to the BamHI site.
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Vector typeMammalian 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)DH10B
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameCer(1-158)-gamma-1
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Alt nameGNGT1
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SpeciesB. taurus (bovine); Aequorea victoria
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Insert Size (bp)676
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MutationGgamma-1 was amplified by PCR, which added a BamHI site and a linker sequence (Arg-Ser) to the 5' end and a 3' BglII site. Cloning into the BglII site of Cer(1-158) destroyed the BamHI site.
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Entrez GeneGNGT1
- Promoter CMV
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Tag
/ Fusion Protein
- Cer(1-158) was fused to the amino terminus of Ggamma-1. (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site BglII (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer SP6 (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byMonomeric Cerulean was obtained from David Piston, Vanderbilt University, Nashville, TN. Gamma-1 cDNA was obtained from Janet Robishaw, Weis Center for Research, Geisinger Clinic, Danville, PA.
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
Reference for monomeric Cerulean:
Rizzo MA, Springer GH, Granada B, Piston DW. An improved cyan fluorescent protein variant useful for FRET. Nat Biotechnol. 2004;22(4):445-9. PubMed PMID: 14990965.
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:
Cer(1-158)-gamma-1 in pcDNAI/Amp was a gift from Catherine Berlot (Addgene plasmid # 55623 ; http://n2t.net/addgene:55623 ; RRID:Addgene_55623) -
For your References section:
Analysis of G protein betagamma dimer formation in live cells using multicolor bimolecular fluorescence complementation demonstrates preferences of beta1 for particular gamma subunits. Mervine SM, Yost EA, Sabo JL, Hynes TR, Berlot CH. Mol Pharmacol. 2006 Jul;70(1):194-205. Epub 2006 Apr 26. 10.1124/mol.106.022616 PubMed 16641313