The use of SSCP analysis in the assessment of phytoplasma gene variability

Single-strand conformation polymorphism (SSCP) analysis is a broadly used technique for detecting mutations. The aim of this work was to assess the applicability of SSCP as a new tool for the detection of the molecular variability of uncultivable mollicutes — phytoplasmas. Three phytoplasma regions...

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Bibliographic Details
Published in:Journal of microbiological methods 2008-04, Vol.73 (1), p.69-72
Main Authors: SERUGA MUSI, Martina, KRAJACIC, Miaden, SORIC, Dijana
Format: Article
Language:English
Subjects:
16S rDNA
Bacteriological methods and techniques used in bacteriology
Bacteriology
Base Sequence
Biological and medical sciences
DNA Fingerprinting
DNA, Bacterial - genetics
DNA, Ribosomal - genetics
dnaB
DnaB Helicases - genetics
Fundamental and applied biological sciences. Psychology
Microbiology
Miscellaneous
Molecular Sequence Data
Mollicutes
phylogenetic analysis
Phylogeny
Phytoplasma - classification
Phytoplasma - genetics
Phytoplasma - isolation & purification
Plant Diseases - microbiology
polymorphism
Polymorphism, Single-Stranded Conformational
RNA, Ribosomal, 16S - genetics
Sequence Alignment
Sequence Analysis, DNA
Stolbur
tuf
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Summary:Single-strand conformation polymorphism (SSCP) analysis is a broadly used technique for detecting mutations. The aim of this work was to assess the applicability of SSCP as a new tool for the detection of the molecular variability of uncultivable mollicutes — phytoplasmas. Three phytoplasma regions were investigated: 16S rDNA, tuf gene, and dnaB gene. Fragments amplified by PCR were subjected to SSCP under conditions optimized for each fragment length. In all of the analyzed regions, SSCP revealed the presence of polymorphism undetected by routine RFLP analyses. Reliability of the method was confirmed by the multiple alignments and phylogenetic analyses of representative sequences showing different SSCP profiles.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2008.02.003