Open AccessGenetic transformation of mouse embryos by microinjection of purified DNA.
Author(s) -
Jon W. Gordon,
George Scangos,
D. Plotkin,
James A. Barbosa,
Frank H. Ruddle
Publication year - 1980
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.77.12.7380
Subject(s) - biology , plasmid , pronucleus , recombinant dna , microinjection , microbiology and biotechnology , dna , embryo , pbr322 , homologous chromosome , gene , zygote , southern blot , in vitro recombination , genome , transformation (genetics) , genetics , virology , molecular cloning , embryogenesis , gene expression
A recombinant plasmid composed of segments of herpes simplex virus and simian virus 40 viral DNA inserted into the bacterial plasmid pBR322 was microinjected into pronuclei of fertilized mouse oocytes. The embryos were implanted in the oviducts of pseudopregnant females and allowed to develop to term. DNA from newborn mice was evaluated by the Southern blotting technique for the presence of DNA homologous to the injected plasmid. Two of 78 mice in one series of injections showed clear homology, though the injected sequences had been rearranged. Band intensities from the two positive mice were consistent with the presence of donor DNA in most or all of the cells of the newborns. These results demonstrate that genes can be introduced into the mouse genome by direct insertion into the nuclei of early embryos. This technique affords the opportunity to study problems of gene regulation and cell differentiation in a mammalian system by application of recombinant DNA technology.