Surface display of p75, a Lactobacillus rhamnosus GG derived protein, on Bacillus subtilis spores and its antibacterial activity against Listeria monocytogenes

AMB Express

Table 1 Bacterial strains, plasmids, and primers used in this study

Bacterial strains or plasmids	Description	Reference
Bacterial strains
Escherichia coli DH5α	F−, φ 80dlacZ∆M15, ∆(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(rK−, mK+), phoA, supE44, λ−, thi-1, gyrA96, relA1	Hanahan (1983)
Bacillus subtilis 168	trpC2	Anagnostopoulos and Crawford (1961)
Lactobacillus rhamnosus GG	ATCC 53103	Purchased from ATCC
Listeria monocytogenes	ATCC19115	Purchased from ATCC
Plasmids
pUB19	E. coli-B. subtilis shuttle vector, Ap^r, Km^r	Kang et al. (2019)
pUB19-cotG-p75	Spore display of p75 using the CotG anchor	This study
Primers
CotG-F	5′-CTTCGACGCGTCAGCTGGCACC-3′	This study
CotG-R	5′-GTCCCTGTCGAGCTTCCTCCTCCTCCTTTGTATTTCTTTTTGACTACCCAGCAATTGCCGT-3′	This study
P75-F	5′-AGAAATACAAAGGAGGAGGAGGAAGCTCGACAGGGACGGTCAGTTACAAATCCGG-3′	This study
P75-R	5′-AAGGAAAAAGCGGCCGCTTATAGTGACGGGCGAACCGCAAAGTCAGG-3′	This study

^a ATCC, American type culture collection (Manassas, VA, USA)
^b The bold letters indicate the restriction sites
^c The underlined letters indicate a flexible linker (GGGGS) at the C-terminal of the cotG structural gene