pNZ8149乳酸菌食品級表達載體

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lacF缺陷型食品級篩選標記，nisA啟動子。

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)).

圖譜Map：

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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