Recombinant Production and Purification of Ion Channels

Rapid and efficient generation of recombinant expression plasmids is critical for academic protein science. Creative Bioarray is committed to helping clients achieve efficient, fast, and flexible plasmid construction and recombinant production of ion channels using the cloning technology of homologous recombination in Saccharomyces cerevisiae, accelerating the function research of ion channels.

Background

Recombinant protein production is central to modern biotechnology. Therefore, optimizing protein yield and quality is a critical step. Furthermore, modern protein chemistry mainly relies on the acquisition of recombinant proteins, since only a very limited number of proteins can be purified from natural sources. Ion channels are important membrane proteins whose structural and functional analysis in basic research relies on the ability to produce and purify recombinant ion channels. Obviously, such channel proteins can only be obtained by recombinant gene expression.

Obtaining large numbers of high-quality recombinant ion channels typically requires the creation and screening of a number of plasmid constructs. An ideal high-throughput cloning technique for generating expression plasmids should have the advantage of being flexible and inexpensive to include all desired features in vector design, as well as the ability to introduce specific sequence randomization into the final construct. In vivo homologous recombination in S. cerevisiae is a simple and cost-effective tool, especially for generating expression plasmids, and minimizes the need for in vitro enzymatic reactions for plasmid assembly.

Fig. 1 Workflow for plasmid construction by homologous recombination. (Preisler, 2019)

Our Services

Creative Bioarray focuses on the many potentials of homologous recombination to generate expression plasmids and site-specific mutations. Our research team has established a workflow for homologous recombination in S. cerevisiae, which allows precise and rapid assembly of expression plasmids without introducing any contaminating nucleotide sequences, thereby providing our clients with production services for recombinant ion channels. In addition, we also provide high-quality purification services for these proteins. Our services include:

• Cloning by homologous recombination in S. cerevisiae
• Generation of ion channel GFP fusions
• Sites directed mutagenesis
• Co-expression of two cDNAs from the same plasmid
• Construction of pET-based expression vectors for homologous recombination cloning in yeast
• Purification of recombinant ion channels

Applications

• Large-scale expression and purification of ion channels
• Research on the detailed structures and functions of ion channels
• Sample preparation for cryo-EM imaging
• Drug discovery targeted ion channels

Creative Bioarray has an advanced technology platform to ensure the professional production and purification of recombinant ion channels for clients. We have established protocols for expressing and purifying high-quality sample preparations, paving the way for cryo-EM imaging and high-resolution structure determination. If you need ion channel synthesis services, please feel free to contact us for more information.

Reference

1. Preisler, S. S. recombinant production and purification of ion channels for biomimetic membranes and cryo-em studies : PHD thesis. University of Copenhagen, Department of Biology, 2019.