pBG/-3G:24C (ATCC® 40289)

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

Permits and Restrictions

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Designations pBG/-3G:24C
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.
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.
Distributed in aliquots of 200 ng dried.
Constructed by oligonucleotide-directed mutagenesis from pBGST7 (ATCC 40288). In the transcript, the 3rd position upstream of the 5' splice site has been changed from U to G and the 24th position in the intron has been changed from A to C.
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

Zaug AJ, et al. The Tetrahymena ribozyme acts like an RNA restriction endonuclease. Nature 324: 429-433, 1986. PubMed: 3537808

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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