pNZ8149乳酸菌食品级表达载体-BioVector NTCC质粒载体菌种细胞基因保藏中心

pNZ8149乳酸菌食品级表达载体

BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心www.biovector.net

lacF缺陷型食品级筛选标记，nisA启动子。

pNZ8149 – broad host range vector; lacF for food grade selection for growth on lactose; nisA promoter followed by an NcoI site for translational fusions at the ATG. Contains a terminator after the MCS. Sequence adaptation for cloning in NcoI can result in a change in the second amino acid of a protein (no reference, example: (Mierau et al., 2005)).

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Host strains

All supplied strains are derivatives of L.lactis subsp. cremorisMG1363, a plasmid-free progeny of the dairy starterstrain NCDO712.

The most commonly used host strain isNZ9000. To construct this strain the genes for NisK and NisR were integratedinto the pepN gene of MG1363.The two genes are

transcribed from their own constitutivepromoter.

Lactococcus lactis NZ9000 – pepN::nisRnisK;

Standard host strain for nisin regulated gene expression(NICE). The strain contains the regulatory genes nisR and nisK integrated into the pepN gene (broad range aminopeptidase) (Kuipers et al., 1998; Mierau andKleerebezem, 2005).

Growth and nisin induction forthe NICEsystem inLactococcus lactis

Various media are available for growth oflactococci. The most commonly used

laboratory medium is M17 supplemented withglucose, lactose or other sugars as carbon source and a relevant antibiotic forplasmid selection. The basic ingredients for a largescale medium are 1–3%peptone, 0.5–2% yeast extract, 1–10% carbon source and small amounts ofmagnesium and manganese ions. Individual processes need specific optimizationof the medium components and fermentation conditions.

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