Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni

ÖZCENGİZ, Gülay; OKAY, Sezer

Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni

Sezer OKAY, (Orta Doğu Teknik Üniversitesi, Biyoloji Bölümü, Ankara, Türkiye)

Gülay ÖZCENGİZ (Orta Doğu Teknik Üniversitesi, Biyoloji Bölümü, Ankara, Türkiye)

Turkish Journal of Biology

3 0

Yıl: 2011 Cilt: 35 Sayı: 1 Sayfa Aralığı: 1 - 7 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni

Öz:

Bacillus thuringiensis (Bt) serovar morrisoni 3023 suşuna ait endokitinaz geni (chi3023) polimeraz zincir reaksiyonu (PCR) ile çoğaltılıp E. coli’ye kopyalandı. chi3023 geni (Gen Bankası Erişim Numarası: DQ993175), 676 aminoasit kodlayan 2031 nükleotitten oluşmaktadır. Moleküler ağırlığı 74.5 kDa, pI değeri de 6.0 olarak hesaplandı. Chi3023’ün aminoasit dizisi daha önce dizilenmiş Bt kitinazları ile karşılaştırıldı ve aralarındaki filogenetik ilişki belirlendi. Prematüre Chi3023’ün N-ucundaki 34 aminoasit, tipik sinyal peptidi özelliği göstermektedir. chi3023 geninin homolog ifadesi için E. coli-Bt vektörü pHT315 kullanıldı. chi3023 genini taşıyan rekombinant pHT315BTC’nin Bt serovar morrisoni 3023’e aktarılması sonucunda endokitinaz aktivitesi 23 kat (0,185 U/mg’dan 4,256 U/mg’a) arttı.

Anahtar Kelime: DNA klonlama kitinaz gen teknolojisi Bacillus thuringiensis endokitinaz enzim aktivitesi karakterizasyon gen transferi kökendeşlik

Konular: Biyoloji

Bacillus thuringiensis serovar morrisoni endokitinaz geninin moleküler kopyalanması, karakterize edilmesi ve homolog ifadesi

Öz:

The endochitinase gene (chi3023) of Bacillus thuringiensis (Bt) serovar morrisoni strain 3023 was amplified via polymerase chain reaction (PCR) and cloned in Escherichia coli. The ORF of chi3023 (GenBank Accession Number: DQ993175) consists of 2031 nucleotides encoding a 676-residue protein with a calculated molecular mass of 74.5 kDa and a pI value of 6.0. The amino acid sequence of Chi3023 was compared with previously sequenced Bt chitinases and the phylogenetic relationships among them were determined. The deduced N-terminal 34 amino acids of the premature Chi3023 exhibited a typical signal peptide. The E. coli-Bt shuttle vector pHT315 was used for homologous expression of chi3023. Introduction of recombinant pHT315BTC, carrying chi3023 into Bt serovar morrisoni 3023, resulted in a 23-fold increase in endochitinase activity (0.185 U/mg versus 4.256 U/mg).

Anahtar Kelime: endochitinase enzyme activity characterization gene transfer homology DNA cloning chitinase gene technology Bacillus thuringiensis

Konular: Biyoloji

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	OKAY S, ÖZCENGİZ G (2011). Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. , 1 - 7.
Chicago	OKAY Sezer,ÖZCENGİZ Gülay Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. (2011): 1 - 7.
MLA	OKAY Sezer,ÖZCENGİZ Gülay Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. , 2011, ss.1 - 7.
AMA	OKAY S,ÖZCENGİZ G Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. . 2011; 1 - 7.
Vancouver	OKAY S,ÖZCENGİZ G Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. . 2011; 1 - 7.
IEEE	OKAY S,ÖZCENGİZ G "Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni." , ss.1 - 7, 2011.
ISNAD	OKAY, Sezer - ÖZCENGİZ, Gülay. "Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni". (2011), 1-7.

APA	OKAY S, ÖZCENGİZ G (2011). Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. Turkish Journal of Biology, 35(1), 1 - 7.
Chicago	OKAY Sezer,ÖZCENGİZ Gülay Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. Turkish Journal of Biology 35, no.1 (2011): 1 - 7.
MLA	OKAY Sezer,ÖZCENGİZ Gülay Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. Turkish Journal of Biology, vol.35, no.1, 2011, ss.1 - 7.
AMA	OKAY S,ÖZCENGİZ G Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. Turkish Journal of Biology. 2011; 35(1): 1 - 7.
Vancouver	OKAY S,ÖZCENGİZ G Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni. Turkish Journal of Biology. 2011; 35(1): 1 - 7.
IEEE	OKAY S,ÖZCENGİZ G "Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni." Turkish Journal of Biology, 35, ss.1 - 7, 2011.
ISNAD	OKAY, Sezer - ÖZCENGİZ, Gülay. "Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni". Turkish Journal of Biology 35/1 (2011), 1-7.