pYES2.1 TOPO™ TA Yeast Expression Kit
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Invitrogen™

pYES2.1 TOPO™ TA Yeast Expression Kit

The pYES2.1 TOPO TA Expression Kit offers direct cloning of a Taq-amplified PCR product into a Saccharomyces cerevisiae expression vector.Read more
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Catalog number K415001
Price (USD)
1,254.00
Each
Add to cart
Price (USD)
1,254.00
Each
Add to cart
The pYES2.1 TOPO TA Expression Kit offers direct cloning of a Taq-amplified PCR product into a Saccharomyces cerevisiae expression vector. The kit uses the linearized, topoisomerase I-activated pYES2.1/V5-His-TOPO ¤ vector for 5-minute cloning on your bench top and produces >85% recombinants. The pYES2.1/V5- His-TOPO™ vector features:

• 2μ origin of replication for high-copy maintenance gene for auxotrophic selection on economical minimal media
• C-terminal V5 epitope and polyhistidine (6xHis) tags for efficient detection and purification
WARNING: Cancer and Reproductive Harm – www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
Specifications
PromoterGAL1
Product TypeTA Yeast Expression Kit
Antibiotic Resistance BacterialAmpicillin (AmpR)
Protein TagHis Tag (6x), V5 Epitope Tag
Cloning MethodTOPO-TA
Quantity20 reactions
VectorpYES, TOPO-TA Cloning Vectors
Product LineTOPO™, YES™
Unit SizeEach
Contents & Storage
Each pYES2.1 TOPO™ TA Expression Kit contains two boxes. The pYES2.1 TOPO™ TA box contains all of the reagents necessary for ligation, including 200 ng of topoisomerase I-activated pYES2.1/V5-His-TOPO™ vector, sterile water, dNTPs, 10X PCR buffer, salt solution, control template and primers, GAL1 Forward and V5 C-term Reverse Primers for sequencing or PCR screening, and an expression control plasmid. Store at -20°C. The One Shot™ Box contains all of the reagents necessary for transformation, including twenty-one 50 μl aliquots of chemically competent TOP10F´ E. coli, S.O.C. medium, and supercoiled plasmid control pUC19. Store at -80°C. All components are guaranteed stable at the indicated temperature for 6 months when properly stored.

Frequently asked questions (FAQs)

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

What are the different kinds of media used for culturing Pichia pastoris and S. cerevisiae?

Following are the rich and minimal media used for culturing Pichia pastoris and S. cerevisiae:

Rich Media:
S. cerevisiae and Pichia pastoris
YPD (YEPD): yeast extract, peptone, and dextrose
YPDS: yeast extract, peptone, dextrose, and sorbitol

Pichia pastoris only
BMGY: buffered glycerol-complex medium
BMMY: buffered methanol-complex medium

Minimal Media (also known as drop-out media):
S. cerevisiae
SC (SD): Synthetic complete (YNB, dextrose (or raffinose or galactose), and amino acids)

Pichia pastoris
MGY: minimal glycerol medium
MD: minimal dextrose
MM: minimal methanol
BMGH: buffered minimal glycerol
BMMH: buffered minimal methanol

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Will the Saccharomyces cerevisiae alpha-factor secretion signal be recognized by Schizosaccharomyces pombe?

S. pombe cannot generate P factor when P factor is replaced for alpha in the alpha factor gene. It can, however, produce alpha factor when alpha is replaced for P in the P factor gene. This is negative evidence that S. pombe can process its own mating factor cleavage sites, but not all the cleavage sites of the S. cerevisiae alpha factor. It is better to use a more generic signal sequence (rather than a pre- pro- signal sequence such as alpha). If it is necessary to go the pre- pro- route, it is better to use the S. pombe P factor leader rather than the S. cerevisiae alpha leader.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Do you offer a TOPO-adapted yeast expression vector?

Yes, we do offer the pYES2.1/V5-His-TOPO vector, which is part of the pYES2.1 TOPO TA Expression Kit (Cat. No. K415001), for the direct cloning of Taq polymerase-amplified PCR products and regulated expression in Saccharomyces cerevisiae using galactose.

View all pYES2.1 TOPO™ TA Yeast Expression Kit FAQs