The Transformation of Alkalophilic Xylanase Gene From Bacillus Sp. Origins To Gr(+) Bacteria by Using the Protoplast Transformation Method and Searching Its Expression Level
Abstract
ÖZET PROTOPLAST TRANSFORMASYON METODU KULLANILARAK ALKALİFİLİK Bacillus sp. KÖKENLİ KSİLANAZ GENİNİN Gr (+) BAKTERİLERE TRANSFERİ ve EKSPRESYON DÜZEYİNİN ARAŞTIRILMASI KAVAL, Elif Yüksek Lisans Tezi, Biyoloji Anabilim Dalı Tez Danışmanı: Yrd. Doç. Dr. Nursel DOSTBİL Temmuz 2002, 52 Sayfa Bu çalışmada Van gölü çevresindeki suyu çekilmiş alanlardan alınan topraklardan alkalifilik Bacillus sp. izolasyonu yapılmıştır. Bu suşlar pH 9-11 aralıklarındaki besi ortamlarında üreme karakteri göstermiştir. Alkalifilik Bacillus sp. suşlarında plasmide bağlı ksilanaz üretme yeteneği gösteren geni araştırılmıştır. Denenen suşlarda ksilanaz üretme yeteneği pozitif olanlar acridine orange ile muamele edilerek plasmid eliminasyonu yapılmıştır. Bu aktiviteyi sağlayan plasmide bağlı gen Bacillus sp. X151 suşunda bulunmuştur. Bu sustan plasmid izolasyonu yapılarak bu plasmidlerin protoplast transformasyon tekniği kullanılarak Bacillus cereus P109 suşuna gen aktarımı yapılmış ve ekspresyon düzeyi araştırılmıştır. Anahtar Kelimeler: Alkalifilik Bacillus sp., Ksilanaz, Protoplast Transformasyonu, Ekspresyon
ABSRACT THE TRANSFORMATION OF ALKALOPHILIC XYLANASE GENE FROM Bacillus sp. ORIGINS TO Gr (+) BACTERIA BY USING THE PROTOPLAST TRANSFORMATION METHOD AND SEARCHING ITS EXPRESSION LEVEL KAVAL, Elif MSc, Biological Science Supervisor: Asst. Prof. Dr. Nursel DOSTBİL July, 2002, 52 Pages Alkalophilic Bacillus sp. isolation was made from the soil which was taken from the land around Van Gölü in this research. Strains had fertility characters in the medium at pH 9-11. Xylanase gene from plasmid origins was searched in alkalophilic Bacillus sp. strains. The strains which had positive result in xylanase synthesis were selected and the plasmid of these strains were eliminated by using Acridine Orange. The gene which was belonged to the plasmid providing this activity was found in Bacillus sp. XI 51 strain. By making plasmid isolation from this strain and with the protoplast transformation technique, these plasmids transferred to Bacillus cereus PI 09 strain and expression level was investigated. Key Words: Alkalophilic Bacillus sp., Xylanase, Protoplast Transformation, Expression m
ABSRACT THE TRANSFORMATION OF ALKALOPHILIC XYLANASE GENE FROM Bacillus sp. ORIGINS TO Gr (+) BACTERIA BY USING THE PROTOPLAST TRANSFORMATION METHOD AND SEARCHING ITS EXPRESSION LEVEL KAVAL, Elif MSc, Biological Science Supervisor: Asst. Prof. Dr. Nursel DOSTBİL July, 2002, 52 Pages Alkalophilic Bacillus sp. isolation was made from the soil which was taken from the land around Van Gölü in this research. Strains had fertility characters in the medium at pH 9-11. Xylanase gene from plasmid origins was searched in alkalophilic Bacillus sp. strains. The strains which had positive result in xylanase synthesis were selected and the plasmid of these strains were eliminated by using Acridine Orange. The gene which was belonged to the plasmid providing this activity was found in Bacillus sp. XI 51 strain. By making plasmid isolation from this strain and with the protoplast transformation technique, these plasmids transferred to Bacillus cereus PI 09 strain and expression level was investigated. Key Words: Alkalophilic Bacillus sp., Xylanase, Protoplast Transformation, Expression m
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Keywords
Biyoloji, Bacillus Sphaericus, Bakteriler, Ekspresyon, Genler, Ksilanaz, Protoplast Transformasyonu, Biology, Bacillus Sphaericus, Bacteria, Expression, Genes, Xylanese, Protoplast Transformation
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64