Genetic engineering techniques for lactic acid bacteria: construction of a stable shuttle vector and expression vector for β-glucuronidase

Shiao-Ming Chang; Tsong-Rong Yan

doi:10.1007/s10529-013-1363-7

Genetic engineering techniques for lactic acid bacteria: construction of a stable shuttle vector and expression vector for β-glucuronidase

Biotechnol Lett. 2014 Feb;36(2):327-35. doi: 10.1007/s10529-013-1363-7. Epub 2013 Oct 8.

Authors

Shiao-Ming Chang¹, Tsong-Rong Yan

Affiliation

¹ Institute of Bioengineering, Tatung University, Taipei, 104, Taiwan.

PMID: 24101246
DOI: 10.1007/s10529-013-1363-7

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Gene Expression
Genetic Engineering / methods*
Genetic Vectors*
Genomic Instability
Lactobacillus / genetics*
Lactococcus lactis / genetics*
Streptococcus thermophilus / genetics*
Transformation, Bacterial