Shop All Algae & Cyanobacteria Expression Kits

GeneArt™ Cryopreservation Kit for Algae (Invitrogen™)

The GeneArt® Cryopreservation Kit for Algae is used to preserve algal strains and clones for storage at -80°C for years, thus eliminating liquid nitrogen storage and continuous cultures as a way to maintain your clones. Cryopreservation is the optimal choice for preservation and long-term storage of algae, since it minimizes genetic drift, facilitates strain and clone exchange between labs, and helps reduce maintenance labor and costs. Simply grow your cells in the presence of Cryopreservation Reagent A for 2–5 days, harvest, resuspend in Cryopreservation Reagent B, and freeze a small aliquot in one of two recommended freezing containers* at -80°C. The GeneArt® Cryopreservation Kit for Algae has been used to preserve Chlamydomonas and Chlorella strains with 100% resuscitation after thawing.

• Preserve Chlamydomonas and Chlorella strains and clones for -80°C storage
• Eliminate liquid nitrogen storage
• Minimize contamination and genetic drift that typically occur in continuous cultures
• Ship algal clones on dry ice for exchange between labs

To date, most available methods for algae cell preservation have required liquid nitrogen storage, but this method is inconvenient and expensive. Alternatively, researchers have relied on continuous cultures to maintain algal clones, but this often leads to contamination and genetic drift and is not amenable to shipping. The GeneArt® Cryopreservation Kit for Algae provides a way to freeze algae cells at -80°C for long-term storage or shipment on dry ice with minimized loss in viability.

*We have used the following freezing containers with this product:
• Thermo Scientific™ Mr. Frosty® Freezing Container
• Biocision™ CoolCell™ FTS30 Cell Freezing Container

GeneArt™ Synechococcus Protein Expression Vector (Invitrogen™)

The GeneArt™ Synechococcus Protein Expression Vector is our second-generation Synechococcus protein expression vector optimized for relative high-level expression in algae and with dual protein tags for detection and/or purification of your gene of interest. The kit includes expression vector and easy-to-follow protocols. Our Gibco™ BG-11 Media, sold separately, is optimized for the growth and maintenance of select Cyanobacteria including Synechococcus elongatus, available from the ATCC collection.

• Express >10% total soluble protein of your gene of interest
• Detect and purify your gene of interest with 6His-TEV and/or V5-His epitope tags
• Compatible with seamless assembly for creation of constructs

A specific decided expression vector
The GeneArt Synechococcus Protein Expression Vector is designed to provide constitutive expression in Synechococcus elongatus. Expression is controlled by promoter psbA1, a ribosome binding site (RBS), and optional initiation and stop codons depending on gene location. The vector provides purification and detection tags at the N- and C-termini: an N-terminal polyhistidine tag with a TEV recognition site for optional cleavage and a C-terminal V5 epitope followed by a polyhistidine tag. Two MCS sites provide the flexibility to attach either, both, or no tags toyour protein. Our pSyn_6 vector is also compatible with seamless cloning, an optional cloning method that results in no extra sequences in your final construct.

Features of the vector include:
• Strong constitutive promoter, psbA1, for robust expression of recombinant gene of interest
• Ribosome binding site (RBS) for improved expression of some genes
• Targeted integration of your gene of interest into the Synechococcus elongatus genome
• N-terminal 6His-TEV and C-terminal V5-6His epitope tags
• Spectinomycin-resistance gene for selection in Synechococcus
• Flexible, multiple cloning site vector

>80% integration efficiency
The transformation of Synechococcus elongatus PCC7942 relies on homologous recombination between the cell’s chromosome and exogenous DNA that is not autonomously replicating and contains sequences homologous to the chromosome. The location of integration into the chromosome (neutral site, NS1) was developed as a cloning locus (Clerico et al., 2007), since it can be disrupted without any aberrant phenotype, thus allowing recombination of ectopic sequences. When transformed with vectors containing an antibiotic resistance cassette and neutral site sequences, a double homologous recombination event occurs between the neutral site vector and the Synechococcus elongatus chromosome. The selective marker (spectinomycin) and the gene of interest driven by a promoter are inserted into the neutral site and the vector backbone (pUC) is lost, allowing the expression of recombinant genes in Synechococcus elongatus PCC 7942.

Gibco BG-11 Media—optimized for cyanobacteria
Gibco BG-11 Media, available separately, is optimized for the growth and maintenance of select Cyanobacteria including Synechococcus elongatus. 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 Cyanobacteria culture results in better overall efficiency and more robust data.

MAX Efficiency™ Transformation Reagent for Algae (Invitrogen™)

MAX Efficiency® Transformation Reagent for Algae is a buffer that enhances transformation efficiency for multiple strains of Chlamydomonas species. One of the biggest hurdles in research and development with Chlamydomonas has been the introduction of exogenous DNA into Chlamydomonas strains due to rigid cell walls. Methods such as glass bead agitation, electroporation, and microparticle bombardment are available but often result in low transformation efficiency.

MAX Efficiency® Transformation Reagent for Algae increases the permeability of the Chlamydomonas cell wall and facilitates increased delivery of DNA into the cell nucleus by electroporation. Simply grow the Chlamydomonas cells to early log phase, harvest by centrifugation, and wash twice with MAX Efficiency® Transformation Reagent prior to electroporation. To date, we have seen >200-fold increase in transformation efficiency over previously recommended conditions in 10 different Chlamydomonas strains, including wild type and mutants, using circular or linear DNA, as well as PCR fragments.

• Obtain >1000 transformants/µg DNA for cell wall(+) strains
• Obtain >100 transformants/µg DNA for cell wall(-) strains
• Transform circular or linear DNA or PCR fragments

GeneArt™ Chlamydomonas Protein Expression Vector (Invitrogen™)

The GeneArt™ Chlamydomonas Protein Expression Vector is our second-generation Chlamydomonas protein expression vector for transgene expression from the nuclear genome of eukaryotic green alga Chlamydomonas reinhardtii 137c. It is optimized for relative high-level expression, provides selection against gene silencing, and offers dual protein tags for detection and/or purification of your gene of interest. The kit includes expression vector and easy-to-follow protocols. Our Gibco™ TAP Media, offered separately, is optimized for the growth and maintenance of Chlamydomonas. Chlamydomonas reinhardtii 137c is available from the Chlamydomonas Resource Center.

• Express up to 1% total soluble protein of your gene of interest
• Select against gene silencing even over multiple passages
• Detect and purify your gene of interest with 6His-TEV and/or V5-His epitope tags
• Compatible with seamless assembly for creation of your constructs
• Enables you to receive reliable results with exceptional strain viability and purity

Chlamydomonas pChlamy_4 vector
Transgene expression from the Chlamydomonas nuclear genome offers several advantages over chloroplast expression, such as post-translational modifications and protein-targeting and/or secretion. However, expression from the nuclear genome has typically been less than robust, and the molecular mechanism(s) of poor expression are not completely understood. Possible reasons include poor promoters, genome integration position effects, and transgene silencing. The GeneArt Chlamydomonas Protein Expression Vector provides relative high levels of expressed protein and selection against silencing. Our pChlamy_4 vector is also compatible with seamless cloning, an optional cloning method that results in no extra sequences in your final construct.

Features of the vector include:
• Endogenous and constitutive promoter, Hsp70A-RbcS2, is composed of activator Hsp70A and two copies of RBC S2 intron sequence, resulting in increased expression of your gene of interest
• Hsp70A-Rbc S2 hybrid promoter fused to the bleomycin/Zeocin™-resistance gene allows positive transformants to maintain protein expression levels for multiple passages
• Self-cleaving sequence for the 2A peptide from foot-and-mouth-disease-virus (FMDV) mediates proper cleavage between resistance markers and protein-of-interest to yield two discrete proteins
• 3’ UTR for proper transcript termination and possible additional benefits like increased translation efficiency, mRNA stability, and polyadenylation signals
• Dual protein tags 6His-TEV and V5-His epitopes can be fused to both or either ends of your gene of interest or no tag at all

Selection against silencing
In order to circumvent the transgene silencing that often occurs in C. reinhardtii, our new pChlamy_4 vector is designed so that proteins are expressed as transcriptional fusions with the bleomycin/zeocin resistance gene sh-ble (Rasala, et al, 2012). The self-cleaving sequence for the 2A peptide from the foot-and-mouth-disease-virus (FMDV) is placed between the antibiotic resistance gene and the gene of interest. It encodes a short ~20 amino acid sequence that mediates proper cleavage to yield two discrete proteins. With this system we have seen positive transformants maintain high expression levels for much longer than with other systems, even after many passages with or without selection pressure.

MAX efficiency transformation reagent for algae
One of the biggest hurdles in research and development with Chlamydomonas has been the introduction of exogenous DNA into Chlamydomonas strains due to rigid cell walls. Methods such as glass bead agitation, electroporation, and microparticle bombardment are available but often result in low transformation efficiency. MAX Efficiency™ Transformation Reagent for Algae, when used to pretreat the cells prior to electroporation, enhances transformation efficiency for multiple strains of Chlamydomonas species. It increases the permeability of the Chlamydomonas cell wall and facilitates increased delivery of DNA into the cell nucleus by electroporation. To date, we have seen >200 fold increase in transformation efficiency over previously recommended transformation conditions in 10 different Chlamydomonas strains, including wild type and mutants, using circular or linear DNA, as well as PCR fragments.

Gibco TAP Media—optimized for Chlamydomonas
Gibco TAP Media, available separately, 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.