Type

Journal Article

Authors

Douwe van Sinderen
Christian Cambillau
Valérie Campanacci
Jennifer Mahony
François P. Douillard

Subjects

Biochemistry

Topics
recombinant proteins metabolism pharmacology base sequence nisin gene order drug effects gene expression regulation bacterial genetics lactococcus lactis bacteriophages genetic vectors biosynthesis virology

Construction of two Lactococcus lactis expression vectors combining the Gateway and the NIsin Controlled Expression systems. (2011)

Abstract Over the last 10 years, the NIsin Controlled Expression (NICE) system has been extensively used in the food-grade bacterium Lactococcus lactis subsp. cremoris to produce homologous and heterologous proteins for academic and biotechnological purposes. Although various L. lactis molecular tools have been developed, no expression vectors harboring the popular Gateway recombination system are currently available for this widely used cloning host. In this study, we constructed two expression vectors that combine the NICE and the Gateway recombination systems and we tested their applicability by recombining and over-expressing genes encoding structural proteins of lactococcal phages Tuc2009 and TP901-1. Over-expressed phage proteins were analyzed by immunoblotting and purified by His-tag affinity chromatography with protein productions yielding 2.8-3.7 mg/l of culture. This therefore is the first description of L. lactis NICE expression vectors which integrate the Gateway cloning technology and which are suitable for the production of sufficient amounts of proteins to facilitate subsequent structural and functional analyses.
Collections Ireland -> University College Cork -> PubMed

Full list of authors on original publication

Douwe van Sinderen, Christian Cambillau, Valérie Campanacci, Jennifer Mahony, François P. Douillard

Experts in our system

1
Christian Cambillau
University College Cork
Total Publications: 18
 
2
Jennifer Mahony
University College Cork
Total Publications: 87