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

IGN="CENTER">72

<1

pESP-2

GST-Luciferase

64

12.5

pESP-3

GST-Cre

55

<1

pESP-2

* Yields estimated from small-scale preparations.
** MEK1CA is a constitutively active mutant of MEK1.

Conclusions

Stratagenes new pESP-3 yeast expression vector allows C-terminal fusions with GST for one-step purification of full-length fusion proteins. The GST tag can be removed by proteolytic cleavage with either thrombin or enterokinase. Stratagenes ESP yeast protein expression and purification system and the pESP-1, pESP-2 and pESP-3 vectors offer researchers the choice for either N-terminal or C-terminal tagging with GST. Because the eukaryotic organism, S. pombe, is used for high-level expression and one-step purification of proteins, the posttranslational modifications of eukaryotic proteins are retained.

Methods

Colony PCR analysis. After transformation, E. coli colony PCR analysis was used to verify insertions in the pESP-3 vector using primers flanking the MCS of the pESP-3 vector. The forward primer, 5-GGCATATCATCAATTGAATAAG-3

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