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A New C-Terminal GST Vector for Protein Production in S. pombe

gene was expressed using the pESP-3 vector. The luciferase-GST fusion protein was purified to near homogeneity with a yield of 12.5 mg/liter (data not shown).

Stratagenes FLAG Western Detection Kit can be used to monitor expression and purification processes. With this kit, recombinant fusion proteins that are tagged with the FLAG epitope at N-terminal, C-terminal and internal positions can be detected. As shown in figure 3, the FLAG epitope contained in the calmodulin-GST fusion protein can be detected before and after purification of the fusion protein.

figure 3

Other pESP Vectors

figure 4

Stratagene previously introduced the pESP-1 and pESP-2 vectors (figure 4), which feature N-terminal tagging with GST for purification.2-4 In addition, the pESP-2 vector offers the option of cloning PCR products through ligation-independent cloning (LIC).12 When the LIC method is used, polypeptides without extraneously added amino acid residues can be obtained after removing the GST tag by enterokinase cleavage.3,4 Both the pESP-1 and pESP-2 vectors are available separately and as components of the ESP system and ESP LIC systems, respectively. The ESP system has been extensively tested for inducible expression of a variety of eukaryotic proteins, including human MEK kinases and rat c-Jun N-terminal kinase (JNK). In these tests, up to 12.5 mg/liter of recombinant proteins has been obtained (table 1).

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