pBGST7 (ATCC® 40288)

Organism: Tetrahymena thermophila Nanney and McCoy  /  Clone Type: Clone  /  Depositors: Univ. Colorado, TR Cech, Univ. Colorado

Permits and Restrictions

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Designations pBGST7
Species Tetrahymena thermophila Nanney and McCoy
Depositors Univ. Colorado, TR Cech, Univ. Colorado
Applications
If linearized with HindIII, the T7 runoff transcript can be used to produce the ribozyme.
Vector
Construct size (kb): 3.400000095367432
Insert
DNA: genomic
Insert lengths(kb): 0.6000000238418579
Gene product: ribozyme having endoribonuclease activity toward recognition sequence CUCU (RNA, ribosomal)
Insert Size (kb) 0.600
Biosafety Level 1
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI/HindIII--2.7, 0.6; EcoRI--3.4; HindIII--3.4.
Distributed in aliquots of 200 ng dried.
If linearized with HindIII, the T7 runoff transcript can be used to produce the ribozyme. The transcript is 716 nt long (274 nt of 5' exon, 413 nt of intervening sequence, and 29 nt of 3' exon).
Lengths of the exon sequences were adjusted so that accurate splicing in E. coli would permit expression of beta-galactosidase activity by alpha-complementation.
The vector was constructed by inserting the T7 promoter sequence into a HaeII site approximately 230 bp upstream of the multiple cloning site.
Classification KINGDOM: Protozoa
References

Cech TR, et al. RNA ribozyme polymerases, dephosphorylases, restriction endoribonucleases and methods. US Patent 4,987,071 dated Jan 22 1991

Been MD, Cech TR. One binding site determines sequence specificity of Tetrahymena pre-rRNA self-splicing, trans-splicing, and RNA enzyme activity. Cell 47: 207-216, 1986. PubMed: 3021333

Product Sheet
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Product Sheet