tailieunhanh - Restriction fragment length polymorphism analysis (RFLP) of some Streptomyces strains from soil

To separate the unknown Streptomyces strains isolated from soil samples, the interspacer regions of 16S-23S rDNA of 14 isolates were amplified with PCR (polymerase chain reaction) and digested with three restriction endonucleases, namely, Bsp143I, HaeIII and MnlI. The restriction patterns were used for RFLP (restriction fragment length polymorphism) analysis. | Turk J Biol 25 (2001) 397-404 © TÜB‹TAK Restriction Fragment Length Polymorphism Analysis (RFLP) of Some Streptomyces Strains from Soil Ekrem ATALAN Yüzüncü Y›l Üniversitesi, Fen Edebiyat Fakültesi, Biyoloji Bölümü, Van - TURKEY Received: Abstract: To separate the unknown Streptomyces strains isolated from soil samples, the interspacer regions of 16S-23S rDNA of 14 isolates were amplified with PCR (polymerase chain reaction) and digested with three restriction endonucleases, namely, Bsp143I, HaeIII and MnlI. The restriction patterns were used for RFLP (restriction fragment length polymorphism) analysis. A dendrogram were constructed using the unweighed pair group method using arithmetic averages algorithm (UPGMA) after analysis restriction patterns. Five RFLP groups were obtained and one test strain was left as a single member group. RFLP profile indicated that unknown strains could be identified with data based on the interspacer region of 16S-23S DNA rapid and quickly. Key Words: Fingerprinting, intergenic rDNA, RFLP, Streptomyces Toprakta ‹zole Edilen Baz› Streptomyces Sufllar›n›n Restriksiyon Fragment Uzunluk Polimorfizm (RFLP) ile Analizi Özet: Toprakta izole edilen ve bilinmeyen Streptomyces sufllar›n› birbirinden ay›rmak için 16S-23S rDNA segmenti polimeraz zincir reaksiyonu (PCR) kullan›larak amplifiye edildi. Elde edilen DNA segmenti Bsp1431, HaeIII ve MnlI restriksiyon enzimleri ile muamele edilerek kesildi. Elde edilen bandlar restriksiyon fragmentlerinin uzunluk polimorfizm (RFLP) analizleri için kullan›ld›. Aritmetik ortalama logaritmas› ve a¤›rl›ks›z çift metodu kullan›larak sonuçlar dendogram fleklinde sunuldu. Test organizmalar› 5 gruba ayr›ld› ve 1 test organizmas› da tek üyeli grup oluflturdu. Bilinmeyen organizmalar 16S-23S rDNA interspacer (ara bölge) segmenti RFLP analizi ile teflhisi yap›ld›. Anahtar Sözcükler: Fingerprinting, intergenik rDNA, RFLP, Streptomyces 397 Restriction Fragment Length Polymorphism Analysis (RFLP) of