畜産物利用1

[V-16-15] Expression of GFP-CC12.1 scFv Fusion Protein in Lactococcus lactis

*Suzuka Oshima¹, Fu Namai¹, Suguru Shigemori¹, Takesi Shimosato¹ (1. Shinshu Univ. IBS.)

For many years, lactic acid bacteria have played a major role in the use of fermented milk. Recently, genetically modified lactic acid bacteria (gmLAB) have attracted attention as microbial therapeutics. Here, we constructed a gmLAB strain producing a green fluorescent protein-CC12.1 single-chain fragment variable fusion protein (GFP-CC12.1 scFv); the fusion protein is expected to serve as a prophylactic and/or therapeutic tool against COVID-19. An open reading frame encoding the scFv was cloned into a lactococcal GFP expression vector, yielding a plasmid. Following sequence confirmation, the plasmid was introduced into Lactococcus lactis NZ9000. Resulting gmLAB was cultured with or without nisin, and the soluble protein fractions were obtained for Western blotting. A GFP-CC12.1 scFv band of the expected size (56.2 kDa) was detected only in the sample prepared from the culture stimulated with nisin. In conclusion, we successfully constructed a gmLAB that produces GFP-CC12.1 scFv.