Streptomyces clavuligerus proteomiği-optimizasyon

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2015

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İşlerel, Tekin Elif
Özcengiz, Gülay
Çırçır Hatıl, Ayça

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Streptomyces clavuligerus, sefamisin C antibiyotiği, bir beta laktamaz inhibitörü olan klavulanik asit ve anti-fungal ve antibakteriyel özelliğe sahip başka klavam bileşikleri üretmektedir. Bu metabolitlerin üretimi ve biyosentezi regülatör metabolizmaya bağlı kompleks regülatör moleküllerin etkileşimini gerektirmektedir (Liras vd., 2008). Streptomyces, dirençlilik, transport, biyosentetik enzimleri kodlayan genlere fiziksel olarak bağlı ya da onlar tarafından regüle edilen regülatör genleri içeren ve böylelikle antibiyoitk biyosentezini düzenleyen gen kümelerine sahiptir (Chater ve Bibb, 1997; Alexander ve Jensen, 1998). Proteomik analizler, Streptomyces’te daha önce karakterize edilmemiş hücresel dinamikleri ve bileşenleri açığa çıkarma potansiyeline sahiptir. Ayrıca proteomik yöntemler kullanılarak belirli koşullar altında ekspresyon seviyelerinde meydana gelen önemli değişimlerin 2D jel elektroforez yöntemi kullanılarak belirlenebileceği ve böylelikle istenen ikincil metabolitlerin üretiminin arttırılmasına yönelik organizmada değişiklikler yapılabileceği gibi (Chaudhary vd., 2013) Streptomyces’te antibiyotik üretimi ve ikincil metabolizma ile bağlantı gösteren morfolojik değişim mekanizmaları açığa çıkarılabilir (Zhou vd., 2011). S.clavuligerus türü için henüz belirlenmiş ve optimize edilmiş bir protein ekstraksiyon metodu ve 2D protein profili çıkarma yöntemi yoktur. Bu çalışmada 4 farklı protein ekstraksiyon yöntemi denenerek ve gerektiği durumlarda modifikasyonlar yapılarak, her ekstraksiyon için 2D jelleri hazırlanmış ve jeller birbirleriyle Delta2D programı ile karşılaştırılarak, protein miktarı ve yoğunluğu, ekstraksiyon yönteminin verimliliği açısından en uygun yöntem olan Fenol Ekstraksiyon yöntemi I (Faurobert vd., 2007) seçilmiştir. Böylelikle ileride yapılacak S.clavuligerus proteom çalışmalarımızın ilk ve en önemli basamağı için optimize bir yöntem belirlemiş bulunmaktayız.

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https://app.trdizin.gov.tr/publication/project/detail/TVRRMU9UUTA
https://hdl.handle.net/11511/49768

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Department of Biology, Project and Design

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Citation Formats

T. E. İşlerel, G. Özcengiz, and A. Çırçır Hatıl, “Streptomyces clavuligerus proteomiği-optimizasyon,” 2015. Accessed: 00, 2020. [Online]. Available: https://app.trdizin.gov.tr/publication/project/detail/TVRRMU9UUTA.