Gateway-based system for optimization of recombinant protein expression in K. lactis

Pavlíček J., Škultétyová Ľ., Daniel P., Bařinka C.

Institute of Biotechnology AS CR, Vídeňská 1083,142 20 Prague 4, Czech Republic

Preparation of sufficient amounts of functional recombinant proteins is typically a major bottleneck in structural biology projects. Here we report the construction and evaluation of a set of Gateway-compatible destination vectors comprising the combination of various N- and C-terminal tags. Recombination of the destination vectors with entry clones of interest results in a series of complementary expression vectors. These can be then used to test and optimize expression levels and solubility of a given protein in the contexts of several expression tags in yeast Kluyveromyces lactis.