hGFAP-Cre
(Plasmid #40591)

• Depositing Lab: Albee Messing
• Publication: Zhuo et al Genesis. 2001 Oct;31(2):85-94.

Backbone

• Vector backbone: pgfa2-lac2
• Backbone manufacturer: Michael Brenner (PMID: 7952266)
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 6600
• Modifications to backbone: LacZ coding region of pgfa2-lac2 replaced by a nuclear-targeted Cre coding sequence.
• Vector type: Mammalian Expression, Bacterial Expression, Mouse Targeting, Cre/Lox

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Cre recombinase
• Alt name: CreNLS
• Alt name: hGFAP-Promoter-Cre-MP-1
• Alt name: Tg95
• Species: H. sapiens (human), M. musculus (mouse); Phage
• Insert Size (bp): 1100
• Mutation: Construct contains a nuclear-targeted Cre recombinase followed by a heterologous mouse intron and polyadenylation signal, all under the control of the gfa2 promoter
• Entrez Gene: GFAP (a.k.a. ALXDRD)
• Promoter: hGFAP promoter
• Tags / Fusion Proteins:
  • NLS (N terminal on insert)
  • MP-1 fragment (mouse protamine 1 intron and polyadenylation signal) (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (destroyed during cloning)
• 3′ cloning site: MluI (destroyed during cloning)
• 5′ sequencing primer: GFAPpro-F (5'-ACTCCTTCATAAAGCCCTCG-3')

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Dr. Brenner, Drs. Raul Torres and Klaus Rajewsky
• Articles Citing this Plasmid:
  • 6 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid encodes a transgene to express the Cre recombinase in astrocytes of transgenic mice. A DNA fragment encoding the Cre recombinase was inserted into an expression cassette containing the gfa2 promoter (Besnard et al., 1991), a 2.2-kb 5' flanking region from the human GFAP (hGFAP) gene. The nuclear targeting signal from the SV40 large T antigen was inserted at the amino terminal end of the coding region, because it reportedly increases the efficiency of DNA excision by the recombinase (Gu et al., 1993). A heterologous intron and polyadenylation signal were provided by sequences from the mouse protamine 1 gene at the 3' end.

The hGFAP-cre transgene was constructed by excising the lacZ coding region from pgfa2-lac2 (Brenner et al., 1994) by BamHI digestion, and replacing it by blunt end ligation with a nuclear-targeted Cre coding sequence. The latter was obtained as a 1.1-kb SalI-MluI fragment from plasmid pTZ19RCreNLS, which was kindly provided by Drs. Raul Torres and Klaus Rajewsky. Both junctions were verified by DNA sequencing.