ABSTRACT
A 2.99 kb mtDNA fragment containing two variable restriction endonuclease sites (EcoRV and XbaI) was subcloned and sequenced from the Mediterranean fruit fly (Ceratitis capitata). This fragment represents approximately one-fifth of the entire mitochondrial sequence. The sequence was aligned with the comparable region from Drosophila yakuba and Anopheles gambiae, resulting in 81.8% and 76.7% identity at the nucleotide level, and 77% and 67.7% identity, respectively, at the amino acid level. The sequenced region includes the complete genes for NADH dehydrogenase 4, NADH dehydrogenase 4L, NADH dehydrogenase 6, and transfer RNAs for proline, threonine and histidine, and part of the genes for NADH dehydrogenase 5 and cytochrome b. Oligonucleotide primers were designed to asymmetrically bracket each of two variable restriction endonuclease sites to allow PCR amplification and subsequent restriction endonuclease analysis of individual fly samples.
Subject(s)
DNA, Mitochondrial/genetics , Diptera/genetics , Polymerase Chain Reaction/methods , Amino Acid Sequence , Animals , Anopheles/genetics , Base Sequence , Drosophila/genetics , Genetic Markers , Haplotypes , Molecular Sequence Data , Polymorphism, Restriction Fragment Length , Sequence Homology, Amino Acid , Sequence Homology, Nucleic AcidABSTRACT
Molecular genetic research on the Mediterranean fruit fly, Ceratitis capitata, will provide tools to permit determination of source populations for new pest infestations. Restriction fragment length polymorphism (RFLP) of mitochondrial DNA provides some interpopulation discrimination. A restriction map, including the informative variable EcoRV and XbaI restriction sites, is constructed for the Mediterranean fruit fly, and several restriction sites are associated with specific gene regions based on polymerase chain reaction-RFLP and sequence analyses. A partial sequence of the mitochondrial 16S ribosomal RNA gene is reported.
