Pulsed-Field Jel Elektroforez (PFGE) Metodu ve Akuatik Organizmalarda Kullanımı

Year 2013, Volume: 9 Issue: 1, 44 - 54, 04.01.2013

Mustafa Türe İlhan Altınok

Abstract

PFGE (Pulsed-Field Jel Electroforez), akuakültürde salgın hastalıklara yol açan izolatların ilişkilendirilmelerinde anahtar rol oynayan bir metottur. Teknik, 10 megabaz çifti (Mbç) kadar geniş molekül ağırlığına sahip DNA parçalarının etkin bir biçimde ayrışmasına olanak sağlamıştır. PFGE, çeşitli patojenlerin epidemiyolojisi, genom büyüklüklerinin belirlenmesi, bakteri kromozomlarının fiziksel ve genetik haritalandırılmalarının gerçekleştirilmesi gibi konularda son yıllarda sıkça kullanılan ve altın standart olarak kabul gören bir metottur. Agaroz içerisindeki bakteri hücresinden elde edilen genomik DNA’nın, uygun enzim ile kesimi sonucu oluşan bant desenlerinin yorumlanması şeklinde gerçekleştirilir. Bu derlemede, PFGE’nin genel özellikleri, çalışma prensibi, tipleri, genel ve akuakültürdeki kullanımından söz edilmiştir.

Keywords

Akuakültür, Epidemiyoloji, PFGE, Kesici enzimler

Method of Pulsed-Field Gel Electroforesis (PFGE) and Its Usage in Aquatic organisms

Abstract

PFGE (Pulsed-Field Gel Electroforesis) played a key role in determining the outbreak-associated isolates in aquaculture. The technique has enable to separate large DNA fragments of up to 10 megabase pairs (Mb) effectively. In recent years PFGE, is a method that comonly used and accepted as the gold standard for epidemiology of various pathogen, determining to genome size and constructing the physical and genetic map of the chromosome of bacteria. PFGE is performed: Genomic DNA is derived from bacterial cell in agarose, cut with the appropriate enzyme and band paterns are described. In this review, the general characteristics of PFGE, prenciples of experiment, types of PFGE, the use of PFGE in general and aquaculture are introduced.

Keywords

Aquaculture, Epidemiology, PFGE, Restriction enzymes

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