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Eukaryotic Cell Growth Protein Expression

 
Yeast Media and Reagents

The over-expression of protein in yeast cells represents a powerful expression system for the source of properly folded and functional eukaryotic protein. Please see CIL Application Note 26 (page 71) for additional information regarding pichia pastoris as a viable expression system to produce isotope-enriched protein for NMR investigations.

Insect Cell Media

The Baculovirus Expression Vector System (BEVS), first introduced in the mid-1980’s, has grown to become the most versatile and widely used eukaryotic vector system employed for the expression of recombinant proteins in cultured insect cells. The BEVS is based on the infection of insect cells with recombinant baculovirus (BV) carrying the gene of interest with the subsequent expression of the corresponding recombinant protein by the insect cells. The most popular insect cell lines used in conjunction with the BEVS are Sf9 (Spodoptera frugiperda) and High FiveTM (Trichopulsia ni ).

CIL is proud to offer BioExpress® 2000, a rich growth media for culturing insect cells.   BioExpress® 2000 is packaged as two components: a solid powder (a proprietary blend of inorganic salts, carbohydrates, and amino acids) and a liquid component (fatty acid solution). Selective amino acid-type labeling is possible with BioExpress® 2000.  

The use of BioExpress® 2000 in the uniform 15N and selective amino acid-type labeling of the catalytic domain of AbI kinase is described in CIL Application Note 14. The use of Experimental Design in the optimization of protein yield using BioExpress® 2000 is exemplified in CIL Application Note 20.

Mammalian Cell Media

There is growing interest in obtaining labeled recombinant protein from mammalian cells because eukaryotic protein expressed in mammalian cells has the greatest probability of being properly folded and functional. CIL offers the only commercially available labeled mammalian media intended for the production of labeled protein suitable for NMR studies. Similar growth characteristics should be obtained using BioExpress® 6000 as with using Dulbecco’s Modified Eagle Medium (DMEM). The amino acid content in BioExpress® 6000 is chemically defined so many different custom labeling strategies may be realized.