       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).

