Strategies for the suppression of peroxidase gene expression in tobacco. I. Designing efficient ribozymes
| Autor: | J M Manners, C L McIntyre |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Molecular Sequence Data
Restriction Mapping Gene Expression Regulation Enzymologic Substrate Specificity Suppression Genetic Cleave Tobacco Gene expression Genetics RNA Catalytic Amino Acid Sequence RNA Messenger Binding site Gene Binding Sites Base Sequence biology Ribozyme RNA Plants Genetically Modified Antisense RNA Plants Toxic Peroxidases Biochemistry Drug Design biology.protein Nucleic Acid Conformation Animal Science and Zoology Agronomy and Crop Science Biotechnology Peroxidase |
| Zdroj: | Transgenic Research. 5:257-262 |
| ISSN: | 1573-9368 0962-8819 |
| DOI: | 10.1007/bf01972879 |
| Popis: | Five short hammerhead ribozymes (Rzs) were constructed and tested, using a range of in vitro reaction conditions, for catalytic activity against the mRNA encoding the lignin-forming peroxidase (TPX) of tobacco. Although all 5 Rzs were shown to be able to cleave the RNA substrate, percentage cleavage varied with pre-denaturation of Rz and substrate, incubation temperature, length of incubation and ribozyme (Rz)-to-substrate ratio. One Rz with two catalytic units and 60 nucleotides of complementary sequence in 3 regions was shown to most efficiently cleave the substrate under all in vitro conditions tested. This ribozyme cleaved better than the two single ribozymes from which it was made. The superior cleaving ability of this Rz was shown to be due to the accessibility of the chosen target site and to the increased length of the hybridizing arms spanning this accessible region of the RNA. |
| Databáze: | OpenAIRE |
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