GeneArt® Synechococcus Protein Expression Kit

Catalog number: A24243

Invitrogen™  Related applications: Protein Expression

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Description

The GeneArt® Synechococcus Protein Expression Kit is our second-generation Synechococcus cloning and expression system. Like the first-generation GeneArt® Synechococcus engineering kits, this expression system employs cyanobacterium Synechococcus elongatus strain PCC 7942, but is optimized for high-level expression and offers dual protein tags for detection and/or purification of your gene of interest. The kit includes frozen cyanobacterial cells, expression vector, One Shot® TOP10 competent E.coli cells, 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.

• 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
• Enables you to get reliable results with exceptional strain viability and purity

A Robust Expression Vector
The MCS vector in the GeneArt® Synechococcus Protein Expression Kit is designed to provide constitutive and robust 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 on your 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

Versatile Cyanobacteria for Study or Bioproduction
The cyanobacterium Synechococcus elongatus strain PCC 7942, is an excellent system for studying photosynthesis, circadian rhythms, nutrient regulation, environmental response, and lipid metabolism. Cyanobacteria, also known as blue-green algae, are a common platform for bioproduction of alcohols and chemicals due to their small genome size and the ease with which it can be genetically manipulated by natural transformation.

>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, 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.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Promoter: psbA1
Cell Line: Synechococcus
Expression: Algae
Product Size: 10 reactions
Detection Method: Gel Electrophoresis
Number of Reactions: 10 Reactions
Protein Tag or Fusion: His Tag (6x)
High Throughput Compatibility: Not High Throughput-Compatible (Manual)

Documents

Manuals & protocols

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