Articles
NUCLEOTIDE SEQUENCE OF APPLE SCAR SKIN VIROID REVERSE TRANSCRIBED IN HOST EXTRACTS AND AMPLIFIED BY THE POLYMERASE CHAIN REACTION
Article number
309_44
Pages
305 – 310
Language
Abstract
Apple scar skin viroid (ASSV) was reverse transcribed in vitro in total nucleic acids from infected tissue and amplified in a reverse transcription-polymerase chain reaction (RT-PCR) assay.
The ASSV cDNA amplified product was cloned and subcloned into the vector pUC9 and pGEM, respectively, and the nucleotide sequence of the insert in each vector was determined.
The sequence is 330 nucleotides in length which can be arranged into the ASSV-specific rod-like structure.
These results indicate that the RT-PCR assay transcribes and amplifies viroid RNA in nucleic acid extracts of infected tissue with very high specificity and fidelity.
Thus, reliable cloning and sequencing analysis of ASSV as well as other viroids can now be performed without viroid purification or the need of large amounts of infected tissue.
The ASSV cDNA amplified product was cloned and subcloned into the vector pUC9 and pGEM, respectively, and the nucleotide sequence of the insert in each vector was determined.
The sequence is 330 nucleotides in length which can be arranged into the ASSV-specific rod-like structure.
These results indicate that the RT-PCR assay transcribes and amplifies viroid RNA in nucleic acid extracts of infected tissue with very high specificity and fidelity.
Thus, reliable cloning and sequencing analysis of ASSV as well as other viroids can now be performed without viroid purification or the need of large amounts of infected tissue.
Authors
X. Yang, A. Hadidi, R.W. Hammond
Keywords
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