pET28a-At-TRL
(Plasmid #32242)

• Depositing Labs: Stan Fields, Jay Hesselberth
• Publication: Schutz et al RNA. 2010 Mar;16(3):621-31. Epub 2010 Jan 14.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5255
• Total vector size (bp): 8638
• Vector type: Bacterial Expression

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: A. thaliana tRNA ligase
• Species: A. thaliana (mustard weed)
• Insert Size (bp): 3300
• Entrez Gene: RNL (a.k.a. AT1G07910, ARABIDOPSIS THALIANA RNA LIGASE, ATRNL, AtRLG1, RNAligase, T6D22.1, T6D22_1, TRNA LIGASE)
• Promoter: T7 RNA Polmyerase
• Tag / Fusion Protein:
  • 6xHistidine (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: T7 Promoter
• 3′ sequencing primer: T7 Terminator

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Hildburg Beier
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

We have found that this protein expresses best in the BL21 (DE3) RIPL cells from Stratagene. Standard BL21 (DE3) strains that lack tRNAs for rare codons do not work well (or at all).