Construction and application of a food-grade expression system for Lactococcus lactis.
A food-grade host/vector expression system for Lactococcus lactis was constructed using alanine racemase gene (alr) as the complementation marker. We obtained an alanine racemase auxotrophic mutant L. lactis NZ9000Δalr by double-crossover recombination using temperature-sensitive integration plasmid pG(+)host9 and a food-grade vector pALR with entirely lactococcal DNA elements, including lactococcal replicon, nisin-inducible promoter PnisA and the alr gene from Lactobacillus casei BL23 as a complementation marker. By using the new food-grade host/vector system, the green fluorescent protein and capsid protein of porcine circovirus type II were successfully overexpressed under the nisin induction. These results indicate that this food-grade host/vector expression system has application potential as an excellent antigen delivery vehicle, and is also suitable for the use in the manufacture of ingredients for the food industry.
State Key Laboratory of Microbial Technology, Shandong University, Jinan, People's Republic of China.
Wenwei
W
Lu
Jian
J
Kong
Wentao
W
Kong
D000411:Alanine Racemase / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D017925:Circovirus / Q000235:genetics / Q000378:metabolism; D019649:Food Industry / Q000379:methods; D005516:Food Microbiology / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007780:Lacticaseibacillus casei / Q000235:genetics / Q000378:metabolism; D013294:Lactococcus lactis / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D009154:Mutation / Q000187:drug effects; D009561:Nisin / Q000494:pharmacology; D010957:Plasmids / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000187:drug effects
D001426:Bacterial Proteins; D036022:Capsid Proteins; D009561:Nisin; D049452:Green Fluorescent Proteins; D000411:Alanine Racemase