Gene expression in Lactococcus lactis

Guchte, Maarten van de; Kok, Jan; Venema, Gerhardus

doi:10.1111/j.1574-6968.1992.tb04958.x

Cited by 105 publications

(14 citation statements)

References 108 publications

(204 reference statements)

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“…Incorporation of the preferred ATG start codon (Pouwels and Leer 1993) did not affect expression levels of CAT, but a single nucleotide change in the sequence between the ribosome binding site (RBS) and the start codon reduced CAT production by a factor six. It is conceivable that this negative effect on production is a result of disturbance of the mRNA structure in the window between the RBS and the start codon (Vellanoweth and Rabinowitz 1992; van de Guchte et al. 1992; Pouwels and Leunissen 1994).…”

Section: Discussionmentioning

confidence: 99%

Use of bacteriocin promoters for gene expression in Lactobacillus plantarum C11

et al. 2004

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Aims: To exploit promoters involved in production of the bacteriocin sakacin P for regulated overexpression of genes in Lactobacillus plantarum C11. Methods and Results: Production of sakacin P by Lact. sakei LTH673 is controlled by a peptide-based quorum sensing system that drives strong, regulated promoters. One of these promoters (P orfX ) was used to establish regulated overexpression of genes encoding chloramphenicol acetyltransferase from Bacillus pumilus, aminopeptidase N from Lactococcus lactis or chitinase B from Serratia marcescens in Lact. plantarum C11, a strain that naturally possesses the regulatory machinery that is necessary for promoter activation. The expression levels obtained were highly dependent on which gene was used and on how the promoter was coupled to this gene. The highest expression levels (14% of total cellular protein) were obtained with the aminopeptidase N gene translationally fused to the regulated promoter. Conclusions: Sakacin promoters permit regulated expression of a variety of genes in Lact. plantarum C11. Significance and Impact of the Study: This study shows the usefulness of regulated bacteriocin promoters for developing new gene expression systems for lactic acid bacteria, in particular lactobacilli.

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Section: Discussionmentioning

confidence: 99%

Use of bacteriocin promoters for gene expression in Lactobacillus plantarum C11

et al. 2004

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“…4). In L. lactis , transcription mostly starts 4–8 bp from the −10 site [14]. Therefore, the 5′‐end of the repB mRNA is probably located at a position near (if not the same as) the 3′‐end of ctRNA.…”

Section: Resultsmentioning

confidence: 99%

Involvement of antisense RNA in replication control of the lactococcal plasmid pND324

Duan

Liu

Supple

et al. 1998

FEMS Microbiology Letters

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pND324 belongs to a family of closely related theta-type plasmids from Lactococcus lactis. An antisense RNA, termed countertranscript (ctRNA), was identified which is complementary to the leader sequence of the mRNA that encodes RepB, a protein essential for plasmid replication. When the synthesis of ctRNA was abolished by site-directed mutagenesis within its promoter region, the mutant replicon showed a 1.8-fold increase in copy number. Similar ctRNA promoter sequences are readily identifiable in 12 other published lactococcal theta-type plasmids, suggesting that they all encode a similar ctRNA-mediated regulatory mechanism.

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“…1996) and useful in other biotechnologically related areas. Transformation systems and cloning vectors for Lactococcus have been developed (Leenhouts and Venema 1993), and gene expression signals have been isolated and characterized (van de Guchte et al. 1992).…”

Section: Discussionmentioning

confidence: 99%

Expression of a thermostable xylanase gene from Bacillus coagulans ST-6 in Lactococcus lactis

Raha

Chang

Sipat

et al. 2006

Lett Appl Microbiol

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Aims: The aim of the study is to evaluate whether xylanase can be used as a potential reporter gene for cloning and expression studies in Lactococcus. Methods and Results: The 750 bp xylanase gene was amplified and subcloned into the unique NheI restriction enzyme site of pMG36e and subsequently transformed into competent Escherichia coli XLI‐blue MRF cells and Lactococcus lactis cells. Bacterial culture containing pMG36e‐Xy has an enzyme activity of 390 μg xylose ml−1 culture 30 min−1, respectively, when compared with 40 μg xylose ml−1 culture 30 min−1 for the negative control (plasmidless strain). Conclusions: The thermostable xylanase gene was successfully expressed in both E. coli and L. lactis. The activity of xylanase can be easily detected by the formation of visible clearing zones around the transformed colonies on Remazol Brilliant Blue‐Xylan (RBB‐Xylan) agar media. However, there were some significant differences in the optimum growth temperature and plasmid stability in the new clones. Significance and Impact of the Study: The constructed reporter vector has the potential to be used as a reporter system for Lactococcus as well as E. coli, and it is an addition to the pool of lactococcal vector systems.

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Gene expression in Lactococcus lactis

Cited by 105 publications

References 108 publications

Use of bacteriocin promoters for gene expression in Lactobacillus plantarum C11

Use of bacteriocin promoters for gene expression in Lactobacillus plantarum C11

Involvement of antisense RNA in replication control of the lactococcal plasmid pND324

Expression of a thermostable xylanase gene from Bacillus coagulans ST-6 in Lactococcus lactis

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