pKLAC1-malE Control Plasmid

Catalog # N3741 was discontinued on January 01, 2013 ;

Product Information
pKLAC1-malE is a control plasmid for the K. lactis Expression Kit (NEB #E1000). The backbone integrative expression vector, pKLAC1, directs high-level expression of recombinant protein from the yeast Kluyveromyces lactis. Vector pKLAC1-malE carries the E. coli malE gene, encoding the maltose binding protein (MBP). pKLAC1-malE can be used to test plasmid transformation, cell culture, and protein production procedures. A typical K. lactis GG799 strain containing integrated pKLAC1-malE will produce > 25 mg/L of MBP (Figure 1).

Figure 1: SDS-polyacrylamide gel electrophoresis of secreted recombinant maltose binding protein (MBP). (A) Detection of secreted MBP by Coomassie staining. Lane 1: Protein Molecular Weight Markers. Lane 2: spent culture medium (15 µl) from wild-type K. lactis cells. Lane 3: spent culture medium (15 µl) from K. lactis cells harboring integrated pKLAC1-malE.(B) Detection of secreted MBP by Western analysis using a polyclonal anti-MBP antibody (NEB #E8031S; 1:2000 dilution) and an anti-rabbit IgG-HRP secondary antibody (1:2000 dilution). Lane 1: culture medium from K. lactis cells (15 µl). Lane 2: culture medium (15 µl) from K. lactis cells containing integrated pKLAC1-malE.

Highlights
- Expression of the fusion gene is under the control of the strong K. lactis P_LAC4-PBI promoter
- K. lactis strain GG799 can be used as an expression host
- Ampicillin resistance for propagation in E. coli (See notes for strain recommendations.)
- Fungal acetamidase selectable marker for integration into the K. lactis genome
Product Categories:

Discontinued Products
- Properties & Usage
  
  Storage Temperature
  -20°C
  Sequence Files
  Fasta GenBank
- Related Products
  Companion Products
  
  K. lactis GG799 Competent Cells
  
  NEB® 5-alpha Competent E. coli (High Efficiency)
  
  NEB® 5-alpha Competent E. coli (Subcloning Efficiency)
  
  NEB® 5-alpha Electrocompetent E. coli
  
  K. lactis Protein Expression Kit
- Product Notes
  NEB 5-alpha Competent E. coli (High Efficiency) (NEB #C2987), NEB 5-alpha Electrocompetent E. coli (NEB #C2989) and NEB 5-alpha Competent E. coli (Subcloning Efficiency) (NEB #C2988) are all recommended for propagation of this control plasmid.
- References
  Colussi, P.A. and Taron, C.H. Unpublished observation
Protocols, Manuals & Usage
- Application Notes
  Coexpression of Multiple Proteins in Kluyveromyces lactis N3741
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
  
  N3741Datasheet-Lot0011210
Tools & Resources
- Web Tools
  DNA Sequences and Maps Tool
FAQs & Troubleshooting
- Tech Tips
  
  Always do the control transformation. This will help mediate any downstream expression issues which may arise.
Quality, Safety & Legal
- Quality Control Assays
  
  Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
  
  N3741Datasheet-Lot0011210
- Legal and Disclaimers
  
  This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
  
  While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
  
  For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
  
  This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
  
  New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
  Licenses
  The vector pKLAC1-malE is a component of an expression system that was developed from basic research at New England Biolabs, Inc. and DSM Biologics Company B.V. The buyer/user has a non-exclusive sublicense to use this system or any component thereof, including vector pKLAC1-malE, for RESEARCH PURPOSES ONLY. A license to use this system for manufacture of clinical grade material or commercial purposes is available from New England Biolabs, Inc., or DSM Biologics Company B.V.