pBI101-pGC1(D1)-GUS
(Plasmid #49513)

• Purpose: Expresses beta-glucuronidase gene under the control of guard cell promoter (pGC1 (D1)) in plant
• Depositing Lab: Julian Schroeder
• Publication: Yang et al Plant Methods. 2008 Feb 19;4:6. doi: 10.1186/1746-4811-4-6.

Backbone

• Vector backbone: pBI101
• Modifications to backbone: To amplify the GC1 (At1g22690) promoter from the Col genomic DNA by PCR, primers YZ27 (5'-CATGCCATGGatttcttgagtagtgattttgaag-3', right before the ATG start codon with NcoI site) and YZ159 (5'-GCGTCGACatggttgcaacagagaggatga-3', 1141 bp upstream of the transcriptional start, D1, with SalI site) were utilized. The PCR product was cloned into pGEM-Teasy vector (Invitrogen, Carlsbad, CA) to create pGEM-T-pGC1. To clone the GC1 promoter into the pBI101 vector, pGEM-T-pGC1 was first cut by NcoI. The sticky end was then filled-in by T4 DNA polymerase (New England BioLabs) to create a blunt end. The pGC1 fragment was then released by SalI digestion. Meanwhile, the destination vector, pBI101, was cut sequentially by SmaI and SalI. The pGC1 fragment was then inserted upstream of the GUS reporter gene in the pBI101 vector to create pBI101-pGC1::GUS construct (simplified as pGC1::GUS).
• Vector type: Plant expression
• Selectable markers: Kanamycin

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Promoter of GC1(D1)
• Alt name: Promoter of GC1 (At1g22690)
• Species: A. thaliana (mustard weed)
• Insert Size (bp): 1140
• GenBank ID: NM_001198136.1 NM_001198136.1
• Entrez Gene: AT1G22690 (a.k.a. AT1G22690, T22J18.14, T22J18_14)
• Promoter: GC1 (At1g22690), 1141 bp upstream of the transcriptional start
• Tag / Fusion Protein:
  • beta-glucuronidase

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (unknown if destroyed)
• 3′ cloning site: SalI (unknown if destroyed)
• 5′ sequencing primer: CATGCCATGGatttcttgagtagtgattttgaag
• 3′ sequencing primer: GCGTCGACatggttgcaacagagaggatga

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Yingzhen Yang

Terms and Licenses

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