Expression of chitinase A (chiA) gene from a local isolate of Serratia marcescens in Coleoptera-specific Bacillus thuringiensis |
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Authors: | Okay S Tefon B E Ozkan M Ozcengiz G |
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Affiliation: | Biological Sciences Department, Middle East Technical University, Ankara, Turkey; Department of Environmental Engineering, Gebze Institute of Technology, Kocaeli, Turkey |
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Abstract: | Aims: The present study focused on cloning and expression of chiA gene from a highly chitinolytic local isolate of Serratia marcescens in an anti-Coleopteran Bacillus thuringiensis and comparison of the characteristics of the native and recombinant ChiAs. Methods and Results: chiA gene from Ser . marcescens was cloned, sequenced and compared with the previously cloned chiA genes. chiA gene was PCR cloned and expressed in anti-Coleopteran B. thuringiensis strain 3023 as verified by Western blot analysis. Specific ChiA activity of the recombinant B. thuringiensis (strain 3023-SCHI) reached its highest level at 21st hour of growth (16·93 U mg−1), which was 5·2- and 1·3-fold higher than that of its parental strain and Ser . marcescens , respectively. Temperature and pH effects on native and recombinant ChiAs were next determined. The recombinant plasmid was quite stable over 240 generations. Conclusions: Serratia marcescens ChiA was heterologously expressed in an anti-Coleopteran B. thuringiensis at levels even higher than that produced by the source organism. Significance and Impact of the Study: Bacillus thuringiensis 3023-SCHI co-expressing anti-Coleopteran Cry3Aa protein and Ser . marcescens chitinase offers a viable alternative to the use of chitinolytic microbes/enzymes in combination with entamopathogenic bacteria for an increased potency because of synergistic interaction between them. |
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Keywords: | Bacillus thuringiensis biocontrol bioinsecticides chitinases Cry3Aa protein Serratia marcescens |
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