Publication: Cloning of a chitinase gene into Bacillus thuringiensis subsp. aizawai for enhanced insecticidal activity
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Issued Date
1997-01-01
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ISSN
00221260
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2-s2.0-0031412589
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of General and Applied Microbiology. Vol.43, No.6 (1997), 341-347
Suggested Citation
Srisurang Tantimavanich, Somsak Pantuwatana, Amaret Bhumiratana, Watanalai Panbangred Cloning of a chitinase gene into Bacillus thuringiensis subsp. aizawai for enhanced insecticidal activity. Journal of General and Applied Microbiology. Vol.43, No.6 (1997), 341-347. doi:10.2323/jgam.43.341 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/17977
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Title
Cloning of a chitinase gene into Bacillus thuringiensis subsp. aizawai for enhanced insecticidal activity
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Abstract
Chitinase from a high producing strain (TP-1) of Bacillus licheniformis was used with B. thuringiensis subsp. aizawai (B.t.a.) in a combined larvicidal assay against the pest, Spodoptera exigua. With 10 mU of this chitinase, the LD50of B.ta. was reduced by 7.6, 13.8 and 15 times on days 3, 5 and 7, respectively when compared to use of B.ta. alone. In addition, a combination of chitinase (10 mU) and B.t.a. at a sub-lethal dose retarded growth and development of S. exigua. In preparation for transformation of B.ta., the TP-1 chitinase gene was cloned in E. coli DH5α and sequenced to reveal a single open reading frame of 1815 bp. This open reading frame encoded for a protein of 604 amino acids and a characteristic signal peptide sequence of 35 amino acids. The gene was subsequently introduced into B.t.a. where it was expressed constitutively. The transformed strain showed slightly improved activity against S. exigua when compared to the non-transformed strain. This was probably due to the low chitinase activity (15 mU/ml) of the transformant, which might be improved by further gene manipulation to overexpress enzyme production.