title: The use of SSCP analysis in the assessment of phytoplasma gene variability.
abstract: 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.
authors.affiliation: Department of Biology, Faculty of Science, University of Zagreb, Croatia. martina@botanic.hr
authors.forename: Martina
authors.initials: M
authors.lastname: Seruga Musić
authors.forename: Mladen
authors.initials: M
authors.lastname: Krajacić
authors.forename: Dijana
authors.initials: D
authors.lastname: Skorić
mesh_terms: D001483:Base Sequence; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D053488:DnaB Helicases / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D045582:Phytoplasma / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D010935:Plant Diseases / Q000382:microbiology; D018807:Polymorphism, Single-Stranded Conformational; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D053488:DnaB Helicases