Abstract
The shuttle vector, pUL6erm, was constructed by using a replicon from pL2, a multiple cloning site, colE1 ori, the ori of Gram-negative bacteria from vector pUC19, and the erythromycin resistance gene from pVA838 as a selection marker. pUL6erm could be transformed easily and maintained stably in Lactococcus lactis, Streptococcus thermophilus, Lactobacillus plantarum and Lactobacillus casei. Transformation assays of pUL6erm indicated that it had a narrow host range. β-Glucuronidase was induced in the presence of 0.3 M NaCl and 50 mM glutamate and expressed at 2.4 U mg−1 with the expression vector (pUL6erm–gadR–GUS) constructed based on pUL6erm carrying β-glucuronidase gene wuth a chloride-inducible (gadR) expression cassette using Pgad as promoter. Therefore, pUL6erm and pUL6erm–gadR–GUS might be a safe and useful genetic tool for the improvement of lactic acid bacteria.
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This work was supported by Grants NSC 90-2214-E-036-005 and NSC 91-2214-E-036-001 from the National Science Council, Republic of China.
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Chang, SM., Yan, TR. Genetic engineering techniques for lactic acid bacteria: construction of a stable shuttle vector and expression vector for β-glucuronidase. Biotechnol Lett 36, 327–335 (2014). https://doi.org/10.1007/s10529-013-1363-7
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DOI: https://doi.org/10.1007/s10529-013-1363-7