Positive-negative selection methods and vectors

Details Positive-negative selection methods and vectors Positive-negative selection methods and vectors

1993-02-04

1995-11-07

Stone, Jacqueline M.

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Modification of viruses

4351721, 4352402, 4353201, 536 231, 536 232, 536 235, 536 236, 536 237, 536 2372, 800 2, 800DIG1, 800DIG2, 935 22, 935 56, 935 70, C12N 1565, C12N 1564, C12N 1509, C12N 1511

Patent

active

054647649

ABSTRACT:
Positive-negative selector (PNS) vectors are provided for modifying a target DNA sequence contained in the genome of a target cell capable of homologous recombination. The vector comprises a first DNA sequence which contains at least one sequence portion which is substantially homologous to a portion of a first region of a target DNA sequence. The vector also includes a second DNA sequence containing at least one sequence portion which is substantially homologous to another portion of a second region of a target DNA sequence. A third DNA sequence is positioned between the first and second DNA sequences and encodes a positive selection marker which when expressed is functional in the target cell in which the vector is used. A fourth DNA sequence encoding a negative selection marker, also functional in the target cell, is positioned 5' to the first or 3' to the second DNA sequence and is substantially incapable of homologous recombination with the target DNA sequence. The invention also includes transformed cells containing at least one predetermined modification of a target DNA sequence contained in the genome of the cell. In addition, the invention includes organisms such as non-human transgenic animals and plants which contain cells having predetermined modifications of a target DNA sequence in the genome of the organism.

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