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Applied and Environmental Microbiology, November 1999, p. 4808-4813, Vol. 65, No. 11
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Inducible Amplification of Gene Copy Number and Heterologous Protein Production in the Yeast Kluyveromyces lactis

Giovanni B. Morlino,1 Lorenza Tizzani,1 Reinhard Fleer,2 Laura Frontali,1 and Michele M. Bianchi1,*

Department of Cell and Developmental Biology, University of Rome "La Sapienza," Rome 00185, Italy,1 Biotechnology Department Rhone-Poulenc Rorer, Vitry 94403, France2

Received 22 March 1999/Accepted 5 August 1999

Heterologous protein production can be doubled by increasing the copy number of the corresponding heterologous gene. We constructed a host-vector system in the yeast Kluyveromyces lactis that was able to induce copy number amplification of pKD1 plasmid-based vectors upon expression of an integrated copy of the plasmid recombinase gene. We increased the production and secretion of two heterologous proteins, glucoamylase from the yeast Arxula adeninivorans and mammalian interleukin-1beta , following gene dosage amplification when the heterologous genes were carried by pKD1-based vectors. The choice of the promoters for expression of the integrated recombinase gene and of the episomal heterologous genes are critical for the mitotic stability of the host-vector system.

* Corresponding author. Mailing address: Department of Cell and Developmental Biology, University of Rome "La Sapienza," p.le Aldo Moro 5, Rome 00185, Italy. Phone: 39 0649912215. Fax: 39 0649912351. E-mail: bianchimic{at}axcasp.caspur.it.

Applied and Environmental Microbiology, November 1999, p. 4808-4813, Vol. 65, No. 11
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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