Biyolüminesans ışıma ve biyolüminesans görüntüleme tekniklerinin moleküler biyoloji araştırmaları bakımından önemi

Canlı bünyesinde meydana gelen reaksiyonlar sonucunda kimyasal enerjiden görünür ışık üretilmesine ve buna bağlı olarak meydana gelen ışımaya biyolüminesans ışıma denir. Biyolminesans gösteren organizmaların sentezledikleri lusiferaz enzimler ve kimyasal dönüşümlerini katalizledikleri ilgili substratların oluşturdukları reaksiyonlar neticesinde biyolüminesans ışıma meydana gelmektedir. Farklı canlı türlerinde çeşitli lusiferaz enzimleri bulunmaktadır. Lusiferaz enzimlerden seçilecek olan birini kodlayan reporter gen, cDNA aracılığıyla herhangi bir proteini kodlayan genle kaynaştırılmak suretiyle, ilgili proteinin lokasyonu veya etkileşimleri in vivo olarak izlenebilmektedir. İlgilenilen virüs, bakteri, parazit ve maya türlerine aktarılan lusiferaz enzim genleri sayesinde, bu türlerin oluşturdukları enfeksiyonların seyir süreçleri izlenebilmektedir. İzleme düzeneği, ilgili denek hayvana lusiferaz geninin aktarılması, hayvana substratın enjekte edilmesi ve CCD kamera (foton-elektron etkileşimli kamera) ile izlemenin yapılması basamaklarından oluşmaktadır. Özellikle protein-protein etkileşim çalışmalarında kullanılan BRET (biyolüminesans ışımaya dayalı rezonans enerji transferi) tekniği ile biyolüminesans ve floresan ışımalar bir arada izlenebilmektedir. Diğer protein saptama/izleme teknikleri ile kıyaslandığında in vivo biyolüminesans görüntüleme denek hayvana girişimde bulunmayı gerektirmeyen, basit, ucuz ve oldukça elverişli bir tekniktir. Bu çalışmada biyolüminesans ışımanın temel prensipleri, biyolüminesans ışıma üreten enzim-substrat sistemleri ve biyolüminesans ışımaya dayalı çeşitli in vivo izleme düzenekleri hakkında genel bilgiler verilmiş ve bu konularla ilgili önemli çalışmaların sonuçları derlenmiştir. 

Bioluminescence radiation and importance of bioluminescence imaging techniques in molecular biology studies

Some unique living organisms produce visible light from chemical energy in a process called bioluminescence. Bioluminescent organisms express luciferase enzymes which catalyze chemical reactions in which luciferases convert their substrates and produce visible light. Different bioluminescent organisms contain different luciferase enzyme/substrate systems. For creating an assay system, genes encoding selected luciferase reporter and any protein, can be fused via cDNA synthesis and then luciferase-fused protein can be traced in living organism or in cell culture, depending on bioluminescence glowing (radiation).  Besides, in a predetermined setup, progression phases of experimentally created infections of any bacteria, virus, parasite or fungus species which transfected with luciferase encoding gene can easily be traced via bioluminescence. Any bioluminescence assay system is composed from three elements: transfer of luciferase gene and injection of its substrate material to animal subject, and acquiring/processing light signals by charge coupled device (CCD) camera.  In bioluminescence resonance energy transfer (BRET) system which is particularly used in protein-protein interaction (PPI) studies, closely positioned or interacting labeled-proteins give both bioluminescent and fluorescent signals. In comparison to other protein-assay techniques, bioluminescence imaging is simple, non-invasive, cost-effective and convenient technique which is promising in terms of finding more usage areas in the future. In this paper, besides a review of important studies focused on the subject, general knowledge about basic principles of bioluminescence, various bioluminescence-creating enzyme-substrate systems and bioluminescence imaging (BLI) modalities were provided. 

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Kaynak Göster

MLA Tunç, E . "Biyolüminesans ışıma ve biyolüminesans görüntüleme tekniklerinin moleküler biyoloji araştırmaları bakımından önemi" . Cukurova Medical Journal 44 (2019 ): 1473-1483 <https://dergipark.org.tr/tr/pub/cumj/issue/42408/535811>