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Improved shuttle vector for expression of chitinase gene in Bacillus thuringiensis
Author(s) -
Lertcanawanichakul M.,
Wiwat C.
Publication year - 2000
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1365-2672.2000.00777.x
Subject(s) - shuttle vector , biology , chitinase , plasmid , xhoi , bacillus thuringiensis , microbiology and biotechnology , amp resistance , tetracycline , bamhi , genetics , recombinant dna , vector (molecular biology) , gene , bacteria , antibiotics
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 Bam HI, Sma I, Pst I, Hin dIII, Sal I, Xho I, Dra II, Apa I and Kpn I 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 Hin dIII 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.