TETRACYCLINE RESISTANT EUKARYOTIC CELLS EXPRESSING AN NADP-REQUIRING OXIDOREDUCTASE

摘要

TETX, a tetracycline degrading enzyme, is provided as a selection marker for eukaryotic cells. Polynucleotide molecules containing tetx and DNA constructs comprising a tetx under the control of a eukaryotic promoter are provided. Additionally, DNA constructs that contain a gene of interest and a tetx, each under the control of an appropriate promoter are provided. Further, methods of expressing tetx in a eukaryotic cell are provided, the method comprising introducing a vector comprising tetx into eukaryotic cells, culturing the eukaryotic cells in the presence of tetracycline, and identifying and isolating the cell that expresses tetx. Furthermore, a method is provided for expressing a gene of interest encoding a protein of interest in a eukaryotic cell by culturing a cell containing a vector comprising a gene of interest and a tetx, each under the control of an appropriate promoter.

主权项

1. A DNA construct comprising a first promoter operably linked to a tetx, wherein tetx is a polynucleotide comprising:
(a) the sequence set forth in any one of SEQ ID NOs: 1-62; (b) a sequence having at least 70% sequence identity to the sequence set forth in any one of SEQ ID NOs: 1-62, wherein the first promoter drives transcription of the tetx when the DNA construct is in a eukaryotic cell.