Publication: Improved shuttle vector for expression of chitinase gene in Bacillus thuringiensis
Issued Date
2000-09-11
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ISSN
02668254
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2-s2.0-0033837188
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Mahidol University
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SCOPUS
Bibliographic Citation
Letters in Applied Microbiology. Vol.31, No.2 (2000), 123-128
Suggested Citation
M. Lertcanawanichakul, C. Wiwat Improved shuttle vector for expression of chitinase gene in Bacillus thuringiensis. Letters in Applied Microbiology. Vol.31, No.2 (2000), 123-128. doi:10.1046/j.1365-2672.2000.00777.x Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/25974
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Title
Improved shuttle vector for expression of chitinase gene in Bacillus thuringiensis
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Abstract
A 6.96-kbp plasmid vector pBCX was constructed from the plasmid pBC16 (4.4 kbp) and a 2.56-kbp fragment of pBluescript II KS. The bifunctional plasmid pBCX conferred ampicillin and tetracycline resistance in Escherichia coli but only tetracycline resistance in Bacillus thuringiensis. It has unique sites for BamHI, SmaI, PstI, HindIII, SalI, XhoI, DraII, ApaI and KpnI derived from pBluescript II KS and was lost at a low rate in B. thuringiensis subsp. israelensis when cultured in Luria-Bertani broth without antibiotic. The chitinase gene from B. circulans number 4.1 (pCHIB1) was subcloned into the HindIII sites of this vector and designated as pBX43 (9.56 kbp). This plasmid produced three times as much chitinase in B. thuringiensis subsp. israelensis strain c4Q272 as pHYB43, which comprises the commercial shuttle vector pHY300PLK plus the chitinase gene.