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Increased production of Coleoptera-specific delta-endotoxin by homologous expression of cry3Aa11 in Bacillus thuringiensis Mm2

The cry3Aa11 gene of Bacillus thuringiensis strain Mm2 (B. thuringiensis Mm2) was PCR-amplified and cloned into E. coli/B. thuringiensis shuttle vector pHT315. The recombinant plasmid carrying the cry3Aa11 gene was introduced into the parental B. thuringiensis Mm2. | A. KURT, G. ÖZCENGİZ Turk J Biol 35 (2011) 585-592 © TÜBİTAK doi:10.3906/biy-0912-15 Increased production of Coleoptera-specific delta-endotoxin by homologous expression of cry3Aa11 in Bacillus thuringiensis Mm2 Aslıhan KURT1,2, Gülay ÖZCENGİZ1 1 Department of Biological Sciences, Middle East Technical University, 06800 Ankara - TURKEY 2 Department of Biology, Yüzüncüyıl University, 65080 Van - TURKEY Received: 14.12.2009 Abstract: The cry3Aa11 gene of Bacillus thuringiensis strain Mm2 (B. thuringiensis Mm2) was PCR-amplified and cloned into E. coli/B. thuringiensis shuttle vector pHT315. The recombinant plasmid carrying the cry3Aa11 gene was introduced into the parental B. thuringiensis Mm2. The toxin production capacities of parental and recombinant strains grown in DSM, containing either 50 mM or 200 mM inorganic phosphate (Pi), were compared by SDS-PAGE and western blot analyses. In comparison to the parental strain, recombinant B. thuringiensis Mm2 produced 4-fold more Cry3Aa11 toxin after 12 h of growth in the presence of 50 mM Pi. However, the 200 mM Pi concentration did not cause a drastic increase in toxin production by the recombinant strain. This study shows that higher toxin production could be obtained from the cultures during the early hours of growth by introducing the multicopy cry3Aa11 gene into the source organism. Key words: Bacillus thuringiensis Mm2, Cry3Aa11 δ-endotoxin, homologous gene expression, cry3Aa11 gene Bacillus thuringiensis Mm2’de cry3Aa11 geninin homolog ifadesi yoluyla Kınkanatlılara-özgü delta-endotoksin üretiminin artırılması Özet: Bacillus thuringiensis Mm2 (B. thuringiensis Mm2) suşuna ait cry3Aa11 geni polimeraz zincir reaksiyonu (PZR) ile çoğaltıldı ve E. coli/B. thuringiensis shuttle vektörüne klonlandı. cry3Aa11 genini taşıyan rekombinant plazmid B. thuringiensis Mm2 suşuna aktarıldı. 50 mM ve 200 mM inorganik fosfat (Pi) içeren DSM besiyerinde üretilen parental ve rekombinant B. thuringiensis suşlarının toksin .