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M9620895.TXT
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1996-02-26
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Document 0895
DOCN M9620895
TI Alteration of hairpin ribozyme specificity utilizing PCR.
DT 9602
AU DeGrandis P; Hampel A; Galasinski S; Borneman J; Siwkowski A; Altschuler
M; Department of Biological Science, Northern Illinois University,;
DeKalb 60115, USA.
SO PCR Methods Appl. 1994 Dec;4(3):139-44. Unique Identifier : AIDSLINE
MED/96000001
AB We have developed a method by which a researcher can quickly alter the
specificity of a trans hairpin ribozyme. Utilizing this PCR method, two
oligonucleotides, and any target vector, new ribozyme template sequences
can be generated without the synthesis of longer oligonucleotides. We
have produced templates with altered specificity for both standard and
modified (larger) ribozymes. After transcription, these ribozymes show
specific cleavage activity with the new substrate beta-glucuronidase
(GUS), and no activity against the original substrate (HIV-1, 5' leader
sequence). Utilizing this technique, it is also possible to produce an
inactive ribozyme that can be used as an antisense control. Applications
of this procedure would provide a rapid and economical system for the
assessment of trans ribozyme activity.
DE Base Sequence *DNA Primers Glucuronidase/BIOSYNTHESIS Human
HIV-1/*GENETICS/METABOLISM Molecular Sequence Data Mutagenesis,
Insertional Plasmids Polymerase Chain Reaction/*METHODS Promoter
Regions (Genetics) RNA, Catalytic/BIOSYNTHESIS/CHEMISTRY/*METABOLISM
Signal Peptides/BIOSYNTHESIS/METABOLISM Support, Non-U.S. Gov't
Transcription, Genetic JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).