Novel use of synthetic oligonucleotide insertion mutants for the study of homologous recombination in mammalian cells.

Abstract:

:Thymidine kinase-deficient mouse L cells have been transformed with plasmid DNAs carrying 8-base-pair Xho I linker insertion mutations in the coding region of the herpes simplex virus type 1 thymidine kinase gene. When the mutant plasmids are introduced individually into LTK- cells, transformation efficiencies are greatly reduced relative to the wild type. However, when two mutant plasmids are cotransferred into the same LTK- recipients, significantly higher frequencies of transformation are observed (30-300 times). Here we demonstrate the usefulness of linker insertions for the study of homologous recombination in detecting the existence of normal thymidine kinase gene sequences (i.e., sequences lacking the insertions after recombination are substantiated by DNA . DNA hybridization). In addition, the frequencies of recombination in the various "crosses" are consistent with the known positions of the mutations.

authors

Shapira G,Stachelek JL,Letsou A,Soodak LK,Liskay RM

doi

10.1073/pnas.80.15.4827

subject

Has Abstract

pub_date

1983-08-01 00:00:00

pages

4827-31

issue

15

eissn

0027-8424

issn

1091-6490

journal_volume

80

pub_type

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