Method of promoting in vitro homologous recombination transfecti

Details Method of promoting in vitro homologous recombination transfecti Method of promoting in vitro homologous recombination transfecti

1993-04-13

1995-11-21

Schwartz, Richard A.

Chemistry: molecular biology and microbiology

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

Modification of viruses

4352402, C12N 1590

Patent

active

054686296

ABSTRACT:
A rapid method for transfection of a cell under physiological conditions suitable to the survival and growth of the cell is disclosed. According to the method, a stable nucleoprotein complex is provided. The nucleoprotein complex comprises a single-stranded DNA sequence in stable combination with RecA protein molecules. Cells to be transformed are cultured in a physiologically suitable medium to which the nucleoprotein complex has been added. As the cells grow and undergo mitosis, the nucleoprotein complex is taken up within some of the cells and becomes integrated into the genome. The method accomplishes transfection without resort to infectious vectors or permeabilization or other manipulation of the cell membrane. According to another object of the invention, a diagnostics method is provided. A directly detectable reporter label or an indirectly detectable ligand is bound to the nucleoprotein complex to provide a DNA probe which then is taken up into the cell and integrates into the cell's genome. Upon appropriate treatment the detectable reporter label can be observed or the ligand can be reacted with a suitable detectable reporter molecule to allow visualization and thereby confirm whether the compliment of the DNA sequence in the probe is substantially present in the genome of the cell.

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