Enhanced heterologous protein productivity by genome reduction in Lactococcus lactis NZ9000

Zhu, Duolong; Fu, Yuxin; Liu, Fulu; Xu, Haijin; Saris, Per Erik Joakim; Qiao, Mingqiang

doi:10.1186/s12934-016-0616-2

Microbial Cell Factories

Table 2 Bacterial strains and plasmids utilized in this study

From: Enhanced heterologous protein productivity by genome reduction in Lactococcus lactis NZ9000

Item	Genotype or phenotype	Reference
Strains
E. coli DH5α	Cloning host; F-φ80lacZ△M15endA1 recA1 endA1 hsdR17 (rK-mK+) supE44 thi-1 gyrA 96 relA1 △ (lacZYA-argF)U169 deoR λ ⁻	[33]
L. monocytogenes WSLC 1019	Animal isolate (ATCC 1916), used as indicator strain	[34]
L. lactis N8	Constitutive strong promoters obtained from this strain	[35]
L. lactis NZ9000 (9 k)	MG1363 pepN::nisRK, the original strain	[36]
L. lactis 9 k-1	The first DNA region L1 (about 9.7 kb) deletion in L. lactis 9 k	This work
L. lactis 9 k-2	The L2 (about 22.5 kb) deletion in L. lactis 9 k-1	This work
L. lactis 9 k-3	The L3 (about 22.2 kb) deletion in L. lactis 9 k-2	This work
L. lactis 9 k-4	The L4 (about 21.6 kb) deletion in L. lactis 9 k-3	This work
L. lactis 9 k-lecC	Em^r, L. lactis 9 k derivative containing pLEB124-lecC	This work
L. lactis 9 k-1-lecC	Em^r, L. lactis 9 k-1 derivative containing pLEB124-lecC	This work
L. lactis 9 k-2-lecC	Em^r, L. lactis 9 k-2 derivative containing pLEB124-lecC	This work
L. lactis 9 k-3-lecC	Em^r, L. lactis 9 k-3 derivative containing pLEB124-lecC	This work
L. lactis 9 k-4-lecC	Em^r, L. lactis 9 k-4 derivative containing pLEB124-lecC	This work
L. lactis 9 k-4-P8/lecC	Em^r, L. lactis 9 k-4 derivative containing pLEB124-P8/lecC	This work
L. lactis 9 k-4-P5/lecC	Em^r, L. lactis 9 k-4 derivative containing pLEB124-P5/lecC	This work
L. lactis 9 k-rfp	Cm^r, L. lactis 9 k derivative containing pNZ8048-rfp	This work
L. lactis 9 k-1-rfp	Cm^r, L. lactis 9 k-1 derivative containing pNZ8048-rfp	This work
L. lactis 9 k-2-rfp	Cm^r, L. lactis 9 k-2 derivative containing pNZ8048-rfp	This work
L. lactis 9 k-3-rfp	Cm^r, L. lactis 9 k-3 derivative containing pNZ8048-rfp	This work
L. lactis 9 k-4-rfp	Cm^r, L. lactis 9 k-4 derivative containing pNZ8048-rfp	This work
Plasmids
pNZ5319	Cm^r, Em^r, used as knock-out vector	[37]
pNZ5319△L1	Cm^r, Em^r, the first DNA region L1 knock-out vector	This work
NZ5319△L2	Cm^r, Em^r, L2 knock-out vector	This work
pNZ5319△L3	Cm^r, Em^r, L3 knock-out vector	This work
pNZ5319△L4	Cm^r, Em^r, L4 knock-out vector	This work
pLEB124	Em^r, containing strong constitutive promoter P45	[38]
pEB690-lecC	Nisin^r, containing the PnisZ+Usp45-lecC+lecI fragment	[23]
pLEB124-lecC	Em^r, pLEB124 derivative containing the P45+PnisZ+Usp45-lecC+lecI fragment	This work
pLEB124-P8/lecC	Em^r, pLEB124 derivative containing the P8+PnisZ+Usp45-lecC+lecI fragment	This work
pLEB124-P5/lecC	Em^r, pLEB124 derivative containing the P5+PnisZ+Usp45-lecC+lecI fragment	This work
pNZ8048	Cm^r	[36]
pET-28a-rfp	Kan^r, containing rfp gene	Aviva systems biology
pNZ8048-rfp	Cm^r, pNZ8048 derivative containing the rfp gene	This work

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