RTS 100 Insect Membrane Kit
- Synthesis of wide range of post-translationally modified eukaryotic proteins
- Expression of membrane proteins without the need for additives
- Yields of up to 50 μg/ml protein, sufficient for functional studies or screening procedures
- Compatible with linear templates and plasmids
- Expression screening in small volumes (>5 μl)
The RTS 100 Insect Membrane Kit, a new eukaryotic cell-free expression system, enables expression of eukaryotic proteins, including membrane proteins, with posttranslational modifications. In contrast to many rabbit-reticulocyte lysate (RRL) -based systems, the insect- cell extract does not require any additives to display full functionality.
The RTS 100 Insect Membrane Kit uses highly productive insect cell lysates obtained from a Spodoptera frugiperda cell line, which contain all translational machinery components (i.e., ribosomes, ribosomal factors, tRNAs, aminoacyl-tRNA synthetases, etc.) required for efficient protein synthesis. In addition, the lysates contain functional organellar membrane fractions, whose activity is required for posttranslational modification of eukaryotic proteins, including membrane proteins. It is a coupled transcription-translation system.
In the in vitro transcription reaction, high-quality capped mRNA is produced using linearized or circular plasmid DNA or PCR products containing a T 7 promoter. In the linked cell-free expression reaction, the capped mRNA is used as template to express active full-length proteins. Using the RTS 100 Insect Membrane Kit, up to 40 gg/ml functionally active posttranslationally modified protein can be synthesized within 3.5 hours.
The RTS plX4.O Insect Vector (cat. no. BR1400901) has been developed for generation of optimal expression templates to be used with the RTS 100 Insect Membrane Kit.