Transformation of Escherichia coli cells by pH 2.3 plasmid DNA treated with psoralens plus near-UV light

1997 ◽  
Vol 37 (1-2) ◽  
pp. 26-30 ◽  
Author(s):  
Rodica Repanovici ◽  
Adriana Plesa ◽  
Gabriela Anton
Keyword(s):  
Escherichia Coli ◽  
Plasmid Dna ◽  
Uv Light ◽  
Escherichia Coli Cells

scholarly journals Effect of polyethylene glycol in plasmid DNA solution on transformation of CaCl2-treated Escherichia coli cells.

1984 ◽  
Vol 48 (3) ◽  
pp. 657-662 ◽  
Author(s):  
Michio HIMENO ◽  
Toshihiro SHIBATA ◽  
Yoshio KAWAHARA ◽  
Yuhji HANAOKA ◽  
Tohru KOMANO
Keyword(s):  
Escherichia Coli ◽  
Polyethylene Glycol ◽  
Plasmid Dna ◽  
Escherichia Coli Cells

scholarly journals Homologous recombination catalyzed by human cell extracts.

1985 ◽  
Vol 5 (4) ◽  
pp. 714-720 ◽  
Author(s):  
R S Kucherlapati ◽  
J Spencer ◽  
P D Moore
Keyword(s):  
Escherichia Coli ◽  
Homologous Recombination ◽  
Gene Conversion ◽  
Human Cell ◽  
Plasmid Dna ◽  
Somatic Cells ◽  
Cell Extracts ◽  
Escherichia Coli Cells ◽  
Enzymatic Machinery

Two plasmids containing noncomplementing and nonreverting deletions in a bacterial phosphotransferase gene conferring resistance to neomycin (Neor) were incubated with human cell extracts, and the mixtures were used to transform recombination-deficient (recA-) Escherichia coli cells. We were able to obtain Neor colonies at a frequency of 2 X 10(-3). This frequency was 100 to 1,000 times higher than that obtained with no extracts. The removal of riboadenosine 5'-triphosphate, Mg2+, or deoxynucleoside triphosphates from the reaction mixture severely reduced the yield of Neor colonies. Examination of plasmid DNA from the Neor colonies revealed that they resulted from gene conversion and reciprocal recombination. On the basis of these results, we conclude that mammalian somatic cells in culture have the enzymatic machinery to catalyze homologous recombination in vitro.

Homologous recombination catalyzed by human cell extracts

1985 ◽  
Vol 5 (4) ◽  
pp. 714-720
Author(s):  
R S Kucherlapati ◽  
J Spencer ◽  
P D Moore
Keyword(s):  
Escherichia Coli ◽  
Homologous Recombination ◽  
Gene Conversion ◽  
Human Cell ◽  
Plasmid Dna ◽  
Somatic Cells ◽  
Cell Extracts ◽  
Escherichia Coli Cells ◽  
Enzymatic Machinery

Two plasmids containing noncomplementing and nonreverting deletions in a bacterial phosphotransferase gene conferring resistance to neomycin (Neor) were incubated with human cell extracts, and the mixtures were used to transform recombination-deficient (recA-) Escherichia coli cells. We were able to obtain Neor colonies at a frequency of 2 X 10(-3). This frequency was 100 to 1,000 times higher than that obtained with no extracts. The removal of riboadenosine 5'-triphosphate, Mg2+, or deoxynucleoside triphosphates from the reaction mixture severely reduced the yield of Neor colonies. Examination of plasmid DNA from the Neor colonies revealed that they resulted from gene conversion and reciprocal recombination. On the basis of these results, we conclude that mammalian somatic cells in culture have the enzymatic machinery to catalyze homologous recombination in vitro.

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