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Large-scale Production of Recombinant RNAs on a Circular Scaffold Using a Viroid-derived System in Escherichia coli
Published on: November 30, 2018
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Cloning and Sequencing of Viroids
1United States Department of Agriculture-Agricultural Research Service (USDA-ARS), Molecular Plant Pathology Laboratory, Beltsville, MD, USA. rose.hammond@usda.gov.
Methods in Molecular Biology (Clifton, N.J.)
|November 30, 2021
Summary
Identifying viroid RNA sequences in plants is crucial for understanding their spread and developing diagnostic tools. New cDNA-based cloning methods accelerate the discovery and characterization of these pathogenic RNAs.
Area of Science:
- Plant pathology
- Molecular biology
- Virology
Background:
- Viroid RNA identification is essential for understanding their distribution in symptomatic and asymptomatic plants.
- Knowledge of viroid distribution aids in developing diagnostic tools and studying viroid biology.
- Recent advances in cDNA-based cloning have expanded the discovery of new viroids.
Purpose of the Study:
- To describe methods for generating complementary DNA (cDNA) copies of viroid RNAs.
- To facilitate the cloning and sequence analysis of viroid RNAs.
Main Methods:
- Complementary DNA (cDNA) synthesis using random primers.
- Complementary DNA (cDNA) synthesis using viroid-specific primers.
- Subsequent cloning and sequence analysis of generated cDNA.
Main Results:
- The described methods enable the generation of cDNA from viroid RNAs.
- These methods support the cloning and sequencing of previously uncharacterized viroids.
- Facilitates comprehensive characterization of viroid populations.
Conclusions:
- Two distinct cDNA synthesis methods are presented for viroid RNA analysis.
- These techniques are vital for expanding knowledge of viroid diversity and distribution.
- Enables improved diagnostics and fundamental research on viroid pathogenesis.
Keywords:
Complementary DNA (cDNA)RT-PCR (reverse transcription-polymerase chain reaction)Reverse transcriptionViroidsMore Related Videos
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