Biological assay of prokaryotic genes in mouse cells following DNA mediated transformation.
Genes coding for leucine biosynthesis in Bacillus subtilis were introduced into mouse LTK- cells by co-transformation with thymidine kinase+ (tk) DNA. Genomic DNA from the tk+ transformants was used to transform competent cultures of different B. subtilis leucine auxotrophs. Each auxotroph was transformed to prototrophy at a similar frequency and the number of leucine gene sequences per transformant genome as deduced by the B. subtilis bioassay strongly correlated with the number estimated by hybridization methods. Tk- subclones were obtained by plating the transformants in 5'-bromodeoxyuridine. One subclone still contained the non-selected leucine gene sequences and could transform auxotrophs of B. subtilis. No deletions or rearrangements in the linkage relationships of the leucine genes occurred in the LTK- cells that inhibited transformation of B. subtilis.
D000818:Animals; D001412:Bacillus subtilis / Q000235:genetics*; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics; D007930:Leucine / Q000235:genetics*; D051379:Mice; D009876:Operon; D013937:Thymidine Kinase / Q000235:genetics; D014170:Transformation, Genetic
D004274:DNA, Recombinant; D004279:DNA, Viral; D013937:Thymidine Kinase; D007930:Leucine