GeneArt® Chlamydomonas TOPO® Engineering Kit

Catalog number: A14260

Invitrogen™  Related applications: Protein Expression

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Description

The GeneArt® Chlamydomonas TOPO® Engineering Kit is the first commercially available genetic modification and expression system for the eukaryotic microalgae, Chlamydomonas reinhardtii. The GeneArt® Chlamydomonas TOPO® Engineering Kit combines TOPO® cloning and expression vectors, frozen cells, simple protocols, and Gibco® TAP media to create the first standardized, complete system for algae research and metabolic engineering in this important model organism. Exceptional purity and our premier TOPO® cloning technology help ensure reliable results and fast ramp up to bioproduction scale.
• Create transformed algal cells typically in just 7-10 days
• Standardize experiments with genetically consistent frozen cells
• Save time required for strain optimization and transition to bioproduction
• Rest easy with excellent strain viability and purity

Versatile and Proven Algal Model Organism
The eukaryotic green algae Chlamydomonas reinhardtii has served as a genetic workhorse and model organism for understanding everything from mechanisms of photosynthesis and nutrient regulated gene expression, to the assembly and function of flagella. Green algae are used as a platform for the production of biofuel and bio-products, due mainly to their rapid growth and ability to convert sunlight and C02 to energy. As C. reinhardtii propagates by vegetative division, the time from initial transformation to product production is significantly reduced relative to plants, requiring as little as six weeks to evaluate production at flask scale, with the potential to scale up to 64,000 liters in four to six weeks.

Shorten the Time from Cloning to Expression
Cells are ready for transformation typically in less than 5 days with straightforward, robust resuscitation and growth protocols. GeneArt® Chlamydomonas pChlamy_3/D-TOPO® Vector facilitates flexible cloning of PCR products for expression in Chlamydomonas reinhardtii. Parallel growth and cloning workflows allow you to obtain transformants typically in just 7-10 days (Fig. 1).

Reliable and Consistent Cell and Vector Performance
Until now, researchers often relied on colleagues and culture collections to obtain cells, vectors, cloning tools, and protocols for working with Chlamydomonas reinhardtii. Cells from these sources often ship on agar slants which could become contaminated and have low survival rates. Additionally, vectors passed down from uncontrolled sources could contain undocumented changes, leading to further wasted time. The GeneArt® Chlamydomonas TOPO® Engineering Kit provides peace of mind with exceptional purity, viability, and cell lot-to-lot consistency. No more genetic drift from ongoing transfer cultures and cloning failures from undocumented sequence changes.

Gibco® TAP Media - Optimized for Chlamydomonas
Gibco® TAP Media, offered separately or in our combo kits, is optimized for the growth and maintenance of Chlamydomonas. The 1X formulation lets you avoid laborious media preparation steps. Award winning bottle design is easier to use in the biosafety cabinet, minimizes the risk of contamination and helps you perform cell culture more consistently. Superior packaging and quality, greater reliability and improved consistency in Chlamydomonas culture results in better overall efficiency and more robust data.

Chlamydomonas reinhardtii TOPO® Cloning Vector
pChlamy_3/D-TOPO® Vector is designed to facilitate rapid, directional TOPO® cloning of blunt-end PCR products for regulated expression in C. reinhardtii. Features of the vector include:
• Hsp70A-RbcS2 constitutive promoter for expression of recombinant gene of interest
• Cop1 3’ UTR for improved expression
• Directional TOPO® cloning site for rapid and efficient directional cloning of a blunt-end PCR product
• Hygromycin resistance gene driven by the beta tubulin promoter for selection in C. reinhardtii
• Ampicillin resistance gene for selection in E. coli
• pUC origin for maintenance in E. coli

For Research Use Only. Not intended for human or animal therapeutic or diagnostic use.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Promoter: Hsp70A-RbcS2
Cell Line: Chlamydomonas
Expression: Algae
Key Function: Algal Engineering
Product Size: 1 kit
Delivery Method: Transformation
Protein Tag or Fusion: Untagged
Selection Agent (Eukaryotic): Hygromycin

Contents & storage

GeneArt® Chlamydomonas TOPO® Engineering Kit contains:



•  10 vials Chlamydomonas reinhardtii — Store at -80 °C

•  1 vial pChlamy_3/D-TOPO® Vector ready for directional TOPO® cloning — Store at -20 °C

•  1 box One Shot® TOP10 Chemically Competent E. coli— Store at -80 °C

•  Protocols for algae resuscitation, growth, transformation, & expression

Documents

Manuals & protocols

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