Multicopy Plasmids Containing the Gene for the Transcriptional Activator LAC9 Are Not Tolerated by K. Lactis Cells

Overview

Journal Curr Genet

Specialty Genetics

Date 1989 Feb 1

PMID 2743432

Citations 5

Authors

K D Breunig

Affiliations

Soon will be listed here.

Abstract

The high copy number 2-microns DNA-like Kluyveromyces plasmid pKD1 was extremely unstable in Kluyveromyces lactis when carrying the gene for the regulatory protein LAC9, a transcriptional activator involved in the induction of the LAC and GAL genes. Transformants of a lac9 mutant strain normally contained rearranged plasmids and all were Lac-, indicating that the LAC9 gene was inactive. Lac+ "revertants" could be obtained from Lac- transformants by selection on lactose plates. In some of these, the pKD1-based plasmid was stably maintained by being integrated into the chromosome of the cell; in others, the disrupted chromosomal gene was restored by a gene conversion event. None of the Lac+ revertants had more than one intact LAC9 gene, an indication that LAC9 overexpression affects cell viability.

Citing Articles

A novel, lactase-based selection and strain improvement strategy for recombinant protein expression in Kluyveromyces lactis.

Krijger J, Baumann J, Wagner M, Schulze K, Reinsch C, Klose T Microb Cell Fact. 2012; 11:112.

PMID: 22905717 PMC: 3520740. DOI: 10.1186/1475-2859-11-112.

Expression of the transcriptional activator LAC9 (KlGAL4) in Kluyveromyces lactis is controlled by autoregulation.

Zachariae W, Breunig K Mol Cell Biol. 1993; 13(5):3058-66.

PMID: 8474461 PMC: 359698. DOI: 10.1128/mcb.13.5.3058-3066.1993.

The yeast Kluyveromyces lactis as an efficient host for heterologous gene expression.

Swinkels B, van Ooyen A, Bonekamp F Antonie Van Leeuwenhoek. 1993; 64(2):187-201.

PMID: 8092859 DOI: 10.1007/BF00873027.

Glucose repression of LAC gene expression in yeast is mediated by the transcriptional activator LAC9.

Breunig K Mol Gen Genet. 1989; 216(2-3):422-7.

PMID: 2501650 DOI: 10.1007/BF00334386.

Coregulation of the Kluyveromyces lactis lactose permease and beta-galactosidase genes is achieved by interaction of multiple LAC9 binding sites in a 2.6 kbp divergent promoter.

Godecke A, Zachariae W, Arvanitidis A, Breunig K Nucleic Acids Res. 1991; 19(19):5351-8.

PMID: 1923819 PMC: 328898. DOI: 10.1093/nar/19.19.5351.

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