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Mitochondrial DNA restriction map for the Caribbean fruit fly,Anastrepha suspensa,and occurrence of mitochondrial DNA diversity within highly inbred colonies
Authors:Heath M  Kuhn D  Schnell R  Olano C
Affiliation:(1) National Germplasm Repository, Subtropical Horticulture Research Station, USDA-ARS, 13601 Old Cutler Rd., Miami, Florida, 33158;(2) Department of Biological Sciences, Florida International University, Miami, Florida
Abstract:A restriction map has been constructed for Anastrepha suspensa mitochondrial DNA. One HaeIII site was found to be polymorphic among individuals in highly inbred colonies and a feral population. Based on mapping information, the polymorphic site was determined to be in the ATPase 6 gene. Primers TK-J-3804 and C3-N-5460 amplified this region. The amplicon was cut by HaeIII in flies of one haplotype and not cut in flies of the other haplotype. From 30 to 43% of the individual flies studied had this additional HaeIII site. After cloning of the ap5200 bp XbaI fragment, the two mitotypes were identified. A 988 base fragment, coding for the entire tRNA-Lys(AAG), tRNA-Asp(GAC), and ATPase 8genes, and a partial ATPase 6gene was sequenced Four silent mutations, including the one at the informative site were located. The HaeIII polymorphism and other sequence differences may prove useful as a diagnostic for identification of the origin of introduced fruitflies.
Keywords:Anastrepha suspensa  Caribbean fruit fly  mitochondrial DNA  mapping  restriction fragment polymorphism
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