Astacus leptodactylus ve Austropotamobius torrentium Türlerinin mtDNA-RFLP Metodu ile Moleküler Teşhisi

Bu çalışmada, canlı türlerinin moleküler olarak teşhisinde yaygın olarak kullanılan mtDNA-RFLP tekniği, morfolojik olarak birbirine benzeyen Austropotamobius torrentium and Astacus leptodactylus kervit türlerinin ayrımında kullanılmıştır. Mitokondrial COI gen bölgesi polimeraz zincir reaksiyonu ile çoğaltılarak 15 farklı endonükleaz enzimi ile kesilmiştir. Bu kesim enzimlerinden üç tanesi (SspI, KpnI, and RsaI) çoğaltılan COI gen bölgesini keserek Austropotamobius torrentium türü için iki parçacık ve üç genotip üretirken, Astacus leptodactylus türü için kesim bölgesi bulamamıştır. Çalışma sonuçları bu üç kesim enziminin mtDNA-RFLP tekniği kullanılarak bu iki kerevit türün ayrımında başarılı bir şekilde kullanılabileceğini göstermiştir

Molecular Identification of Astacus leptodactylus and Austropotamobius torrentium Using mtDNA-RFLP Method

mtDNA-RFLP, a technique commonly used for molecular identification of species, was used to distinguish between morphologically similar crayfish species Austropotamobius torrentium and Astacus leptodactylus in this study. The mitochondrial COI gene was amplified and digested using 15 restriction endonucleases to establish a restriction map. The COI gene sequence contained recognition sites for three of the restriction endonucleases (SspI, KpnI, and RsaI) and produced two fragments and three genotypes for Austropotamobius torrentium. However, the Astacus leptodactylus COI gene did not have restriction sites for these three enzymes. Thus, a mtDNA-RFLP using these three endonucleases can be used to successfully distinguish between these two species of crayfish

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