K. lactis Protease Deficient Competent Cell Sampler

• 产品编号：C1007S      品牌：New England Biolabs       原厂货号：C1007S
• 产品分类：克隆 > 克隆载体和克隆试剂盒 > 细菌菌株和感受态细胞 > 感受态细胞
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描述：

Description:
Chemically competent Kluyveromyces lactis cells are suitable hosts for expressing proteins that have exhibited proteolysis in the parental GG799 background (NEB #C1001S). Proteolysis of secreted heterologous proteins in K. lactis is most often casued by aspartyl poroteases that are present in the secretory pathway. In many cases, a single aspartyl protease causes the majority of the detrimental protein degredation. The Protease Deficient Competent Cell Sampler contains a wild-type GG799 strain, as well as four protease deficient K. lactis strains, each having selectable marker-free deletion of a specific secretory pathway aspartyl protease gene. Each strain can be transformed with any linearized pKLAC-series expression vector, permitting a comparison of heterologous proein quality and yield in each background.

The Sampler contains one transformation each of five K.lactis strains. K. lactis GG799 is a wild-type expression strain. K. lactis YCT389 contains a selectable marker-free deletion of the KLLA0E0398g locus encoding an extracellular aspartyl protease (KlYps1p) that is homologous to cerevisiae Yps1p. K. lactis YCT390 carries a selectable marker-free deletion of the KLLA0F2208g locus encoding an extracellular aspartyl protease (Klyps7p) that is homologous to S.cervisaiae Yps7p. K.lactis strain YCT 569 carries a selectable marker-free deletion of the KLLA0D01507g locus encoding an extracellular aspartyl protease belonging to pfam00026. K.lactis strain YCT598 carries a selectable marker-free deletion of the KLLA0D15917g locus encoding an extracellular aspartyl protease that is homologous to S.cervisaiae Bar1p.

Reagents Supplied:
1 x 0.2 ml/tube of chemically competent K. lactis GG799 cells (NEB #C1001S)
1 x 0.2 ml/tube of chemically competent K. lactis YCT569 cells (NEB #C1003S)
1 x 0.2 ml/tube of chemically competent K. lactis YCT389 cells (NEB #C1004S)
1 x 0.2 ml/tube of chemically competent K. lactis YCT390 cells (NEB #C1005S)
1 x 0.2 ml/tube of chemically competent K. lactis YCT598 cells (NEB #C1006S)
(Store at -80°C)
5 ml of NEB Yeast Transformation Reagent
(Store at 4°C)

 

原厂资料：

注意事项：

General notes:

Storage: Store Competent Cells at -80°C. Once Thawed, Do Not Re-freeze.

Usage notes:

Due to the high transformation efficiency of K. lactis Competent Cells, plating multiple dilutions of the cell mixture is necessary to ensure formation of plates with distinct single colonies. Growth time should not exceed 5 days as small colonies that lack an integrated expression fragment may form.
Plates containing colonies can be stored at 4°C for up to two weeks.
Growth of K. lactis Competent Cells carrying an integrated expression vector for protein expression should be performed in YPGal medium (1% yeast extract, 2% peptone, 2% galactose) supplemented with a final concentration of 38 mM (NH₄)₂SO₄. In the absence of 38 mM (NH₄)₂SO₄, YCT569, YCT389, YCT390 and YCT598 cells will grow slower than the GG799 parent strain (NEB #C1001).