Product Class: Kit

K. lactis Protein Expression Kit

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

Product Introduction

The K. lactis Protein Expression Kit provides an easy method for expressing a gene of interest in the yeast Kluyveromyces lactis. Proteins may be produced intracellularly or secreted using an integrative expression vector (pKLAC2). To achieve protein secretion, a gene of interest is cloned downstream of the K. lactis α-mating factor secretion domain, which is eventually processed in the Golgi and results in secretion of the desired protein.

  • Supplied with competent K. lactis cells (NEB# C1001)
  • Allows cloning and expression of genes toxic to E. coli
  • Rapid high cell density growth results in high yield protein expression 
  • Easy-to-use protocols for those inexperienced with yeast systems
  • No expensive antibiotics or methanol required
  • Attractive commercial sub-licensing
Catalog # Size Concentration
E1000S 1.0 set

Protocols, Manuals & Usage

Protocols

  1. Protein expression using the K. lactis Protein Expression Kit - Linearization of pKLAC2 for integrative transformation of K. lactis.
  2. Protein Expression using the K. lactis Protein Expression Kit - Identification of Multi-copy Integrants
  3. Protein Expression using the K. lactis Protein Expression Kit - Identification of properly integrated cells
  4. Protein Expression using the K. lactis Protein Expression Kit - Simultaneous Expression of Multiple Proteins
  5. Protein expression using the K. lactis Protein Expression Kit - Cloning a PCR fragment into pKLAC2 (E1000).
  6. Protein Expression using the K. lactis Protein Expression Kit - Growth of strains for detection of secreted protein
  7. Protein Expression using the K. lactis Protein Expression Kit - Transformation of K. lactis GG799 cells

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Application Notes

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What systems does NEB offer for protein expression and purification?
  2. I received my kit and don’t see any competent cells or transformation reagent. Where are they?
  3. I’ve expressed my protein, now how do I purify it?
  4. I’ve noticed untransformed K. lactis will grow when streaked on the selection media. Is the selection not working?
  5. Where can I find more detailed FAQs for the K. lactis Protein Expression Kit?
  6. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

Tech Tips

NEBuilder HiFi DNA Assembly Cloning Kit (NEB #E5520) or the Gibson Assembly Cloning Kit (NEB #E5510) are excellent ways to assemble a linear yeast expression cassette. (See "Improved method for assembly of linear yeast expression cassettes using NEBuilder® HiFi DNA Assembly Master Mix" in the Application Notes section under the Other Tools and Resources tab)

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