Drosophila Secretory Expression System

Successful recombination of protein complexes usually requires co-expression of component proteins. This is especially true when component proteins are present as compositional complexes. Based on Drosophila S2 cells, Creative BioMart established a strategy to achieve the large-scale expression of extracellular protein complexes by constructing a stable secretory expression system.

Technology Description

The extracellular domain of extracellular protein or plasma membrane protein plays an important role in various physiological functions. These proteins usually conduct their biological function in the form of homologous or heteromeric protein complexes. At present, the production of protein complexes is usually achieved by using eukaryotic cells to secrete and co-express recombinant proteins.

We use S2 cells here because of their high efficiency of secretory expression. The expression plasmid of the complex component protein was transfected into Drosophila S2 cells together with antibiotic resistance markers, and a stable cell line expressing all components is established.

After transfection of S2 cells, we will detect the expression level of each component and select high expression clones in stable cells. Expression levels are usually analyzed by Western blotting using antibodies against purified tag sequences.

After further passage, the expression of the selected clone is confirmed and the best clone is selected. At this stage, we focus on the characteristics of the clone, such as protein expression level, stability and repeatability. In particular, we focus on the expression level of each component, and preferentially select a clone showing a similar expression level of each component. Then the selected clones are adapted to serum-free medium and large-scale protein production is carried out.

Basic Workflow

Features of S2 Expression System

• The culture condition is simple. The S2 cell line can be easily maintained at room temperature without the need for an incubator with CO 2 supply.
• S2 cells can grow abundantly in the medium, and they also have an effective silencing mechanism (RNAi).
• The S2 cell system supports the constitutive, inducible and transient expression of proteins.
• The inducible vector of the system allows the regulation of gene expression and can be used to produce a large number of proteins for protein crystallography research.
• The S2 cell genome can be modified by a binary expression system.

Related Services

In addition to the secretory expression service, we also offer the following related services.

• Drosophila Genetic Operation Service
• Stable Polyclonal Drosophila S2 Cell Lines Construction Service
• Induction S2 Cell Lines Construction Service
• Drosophila S2 Cell Transient protein production service

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