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Detection of Chrysanthemum stunt viroid (CSV) by nucleic acid spot hybridization

Abstract

Chrysanthemum stunt viroid (CSV) was extracted from Chrysanthemum stunt viroid-infected Chrysanthemum plants. The CSV was then purified to homogenicity from this extract by one cycle of non-denaturating vertical slab gel electrophoresis and was followed by one cycle of denaturating vertical slab gel electrophoresis. No pure circular CSV was obtained. The 3' terminal of purified ASBV was polyadenylated using poly (A) polymerase. The polyadenylated ASBV was used as a template to synthesise radioactively labelled single stranded ASBV cDNA using reverse transcriptase. The radioactively labelled recombinant DNA was hybridised to crude extracts from avocado leaves and chrysanthemum leaves. Autoradiography was employed to detect cDNA-RNA hybrids.
Authors:
Publication Date:
Jan 01, 1985
Product Type:
Book
Reference Number:
AIX-17-019028; EDB-86-054871
Resource Relation:
Other Information: Thesis (M.Sc.)
Subject:
59 BASIC BIOLOGICAL SCIENCES; DNA; HYBRIDIZATION; LABELLING; VIRUSES; PURIFICATION; AUTORADIOGRAPHY; DNA POLYMERASES; ELECTROPHORESIS; PHOSPHORUS 32; PLANT DISEASES; PLANTS; RNA; RNA POLYMERASES; TRANSCRIPTION; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; DAYS LIVING RADIOISOTOPES; ENZYMES; ISOTOPES; LIGHT NUCLEI; MICROORGANISMS; NUCLEI; NUCLEIC ACIDS; NUCLEOTIDYLTRANSFERASES; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; PARASITES; PHOSPHORUS ISOTOPES; PHOSPHORUS-GROUP TRANSFERASES; POLYMERASES; RADIOISOTOPES; TRANSFERASES; 550701* - Microbiology- Tracer Techniques; 550201 - Biochemistry- Tracer Techniques
OSTI ID:
6136518
Research Organizations:
University of the Witwatersrand, Johannesburg (South Africa)
Country of Origin:
South Africa
Language:
English
Availability:
Registrar, University of the Witwatersrand, 1 Jan Smuts Avenue, Johannesburg, 2001, South Africa.
Submitting Site:
INIS
Size:
Pages: 58
Announcement Date:
May 13, 2001

Citation Formats

Bothma, G C. Detection of Chrysanthemum stunt viroid (CSV) by nucleic acid spot hybridization. South Africa: N. p., 1985. Web.
Bothma, G C. Detection of Chrysanthemum stunt viroid (CSV) by nucleic acid spot hybridization. South Africa.
Bothma, G C. 1985. "Detection of Chrysanthemum stunt viroid (CSV) by nucleic acid spot hybridization." South Africa.
@misc{etde_6136518,
title = {Detection of Chrysanthemum stunt viroid (CSV) by nucleic acid spot hybridization}
author = {Bothma, G C}
abstractNote = {Chrysanthemum stunt viroid (CSV) was extracted from Chrysanthemum stunt viroid-infected Chrysanthemum plants. The CSV was then purified to homogenicity from this extract by one cycle of non-denaturating vertical slab gel electrophoresis and was followed by one cycle of denaturating vertical slab gel electrophoresis. No pure circular CSV was obtained. The 3' terminal of purified ASBV was polyadenylated using poly (A) polymerase. The polyadenylated ASBV was used as a template to synthesise radioactively labelled single stranded ASBV cDNA using reverse transcriptase. The radioactively labelled recombinant DNA was hybridised to crude extracts from avocado leaves and chrysanthemum leaves. Autoradiography was employed to detect cDNA-RNA hybrids.}
place = {South Africa}
year = {1985}
month = {Jan}
}