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New Yeast Cloning System for Producing Proteins with Native Amino Acid,,,Sequences

one agarose, the beads were washed and the fusion protein was eluted with 50 mM reduced glutathione. Samples of cells from induced conditions are shown (Figure 3A). Of the eluted GST-calmodulin fusion protein, 20 g was cleaved in a 60-l reaction volume using Stratagenes Enterokinase (figure 3B) at room temperature for 10 or 20 hours.

Acknowledgments

We would like to thank Peter Vaillancourt, Alan Greener and Denise Wyborski for discussions and suggestions and Cherie Dewar for technical assistance.

REFERENCES

  1. Lu, Q., Jerpseth, B., Sanchez, T., Bauer, J.C., and Greener, A. (1997) Strategies 10: 4-6.

  2. Aslanidis, C., and de Jong, P.J. (1990) Nucleic Acids Res. 18: 6069-6074.

  3. Huan, R.S., et al. (1992) Biotechniques 13: 515-518.

  4. Huan, R., and Moss, J. (1992) Gene 112: 37-43.


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