pZJMalpha (ATCC® MBA-140)

Applications: vector for studying DNA-protein interactions tubulin, alpha  /  Depositors: J Morris

Permits and Restrictions

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Designations pZJMalpha
Depositors J Morris
Biosafety Level 1
Vector Information
Size (kb): 6.3710000000000000
Vector: pZJMalpha (plasmid)
Type: expression
Construction: A 656bp fragment of tubulin is cloned into XhoI/HindIII site.
Construct size (kb): 6.4
Features: marker(s): ampR, BleoR
operator: Tet
promoter: T7
replicon: E. coli Ori
MCS: XhoI, XbaI, NdeI, HpaI, HindIII, SfiI
terminator: T7
transcription terminator: Actin3' UTR, Aldolase 3' UTR
Applications
vector for studying DNA-protein interactions tubulin, alpha
Comments
PCR-amplified gene fragments can be ligated in a single step between two opposing T7 promoters. This vector can integrate into a nontranscribed region of the Trypanosoma brucei genome and establish a stable cell line. Tetracycline induction leads to the production of a dsRNA with concomitant degradation of the target mRNA.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Wang Zefeng, et al. Inhibition of Trypanosoma brucei gene expression by RNA interference using an integratable vector with opposing T7 promoters. J. Biol. Chem. 275: 40174-40179, 2000. PubMed: 11013266

Shipped frozen
Restrictions The pZJMalpha was constructed by modifying pLew100 with a cloning site flanked by opposing T7 promoters under the control tetracycline operators. An alpha tubulin gene was inserted between the opposing promoters to prevent the formation of cruciform structures.
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