Detection of point mutations in the chloroplast genome by single-stranded conformation polymorphism analysis.
Single-stranded conformation polymorphism (SSCP) analysis was used to examine the mutations of the chloroplast 16S rRNA locus of streptomycin-resistant mutants in Nicotiana plumbaginifolia. DNA fragments of 121, 517, 968 and 1578 bp, each possessing a known point mutation, were generated by polymerase chain reaction (PCR). The resulting fragments were denatured and separated by nondenaturing polyacrylamide gel electrophoresis. Compared to the patterns of the wild-type DNA fragments, the bands of the single-stranded DNA fragments of 121 and 517 bp with base changes were shifted. However, no pattern variations were detected from the DNA fragments of 968 and 1578 bp generated from both wild-type and mutants.
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Department of Microbiology and Immunology, Chang-Gung Medical College, Taoyuan, Taiwan, Republic of China.
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Liu
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Chang
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D001483:Base Sequence; D002736:Chloroplasts; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D004277:DNA, Single-Stranded; D005240:Feasibility Studies; D016678:Genome; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010940:Plant Proteins / Q000235:genetics; D010947:Plants, Toxic; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics; D014026:Nicotiana / Q000235:genetics
D017931:DNA Primers; D004277:DNA, Single-Stranded; D010940:Plant Proteins; D012336:RNA, Ribosomal, 16S; D012269:Ribosomal Proteins; C034212:ribosomal protein S12