Summary
This communication demonstrates the usefulness of the plamid rescue procedure for recovery of plasmids from transgenic mice. We have microinjected the plasmid pSK1 harbouring the Herpes simplex virus thymidine kinase gene into fertilized mouse oocytes and succeeded in recovering plasmids from newborns by transformation of E. coli either with HindIII cut cellular DNA or with uncut DNA. The majority of the rescued plasmids were indistinguishable from pSK1 by restriction analysis. The rescued plasmids proved to be functionally active in a transient expression assay in mouse Ltk- cells. The pSK1 DNA sequences were inherited by up to 90% of the second generation progeny mice, which is not in agreement with a Mendelian transmission of heterozygous markers integrated into a single site of the chromosome.
These data support the assumption that germ line transmission of non-integrated episomal plasmids can occur.
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References
Birg F, Dulbecco R, Fried M, Kamen R (1979) State and organization of polyoma virus DNA sequences in transformed rat cell lines. J Virol 29:633–641
Birnboim HC, Doly J (1979) A rapid method for the purification of plasmid DNA. Nucl Acids Res 7:1513–1523
Brinster RL, Chen HY, Trumbauer M, Senear AW, Warren R, Palmiter RD (1981) Somatic expression of herpes thymidine kinase in mice following injection of a fusion gene into eggs. Cell 27:223–231
Brinster RL, Ritchie KA, Hammer RE, O'Brien RL, Arp P, Storb U (1983) Expression of a microinjected immunoglobulin gene in the spleen of transgenic. Nature 306:332–336
Campo MS, Cameron IR, Rogers ME (1979) Tandem integration of complete and defective SV40 genomes in mouse-human somatic-cell hybrids. Cell 15:1411–1426
Costantini FD, Lacy E (1981) Introduction of a rabbit β-globin gene into the mouse germ line. Nature 294:92–94
Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995
Folger KR, Wong EA, Wahl G, Capecchi M (1982) Patterns of integration of DNA micro-injected into cultured mammalian cells: evidence for homologous recombination between injected DNA molecules. Mol Cell Biol 2:1372–1387
Gordon JW (1983) Studies of foreign genes transmitted through germ line of transgenic mice. J Exp Zool 228:313–324
Gordon JW, Ruddle FH (1981) Integration and stable germ line transmission of genes injected into mouse pronuclei. Science 214:1244–1246
Gordon JW, Ruddle FH (1982) Germ line transmission in transgenic mice. In: Burger MB, Weber R (eds) Embryonic development, part B: cellular aspects. Liss, New York, pp 112–124
Gordon JW, Ruddle FH (1985) DNA-mediated genetic transformation of mouse embryos and bone marrow — a review. Gene 33:121–136
Gordon JW, Scangos GA, Plotkin DJ, Barbosa JA, Ruddle FH (1980) Genetic transformation of the mouse embryos by microinjection of purified DNA. Proc Natl Acad Sci USA 77:7380–7384
Graham FL, van der Eb A (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467
Hanahan D (1984) Studies on the transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Hanahan D, Lane D, Lipsich L, Wigler M, Botchan M (1980) Characteristics of an SV40-plasmid recombinant and its movement into and out of the genome of a murine cell. Cell 21:127–139
Huttner KM, Scangos GA, Ruddle FH (1979) DNA-mediated gene transfer of a circular plasmid into murine cells. Proc Natl Acad Sci USA 76:5820–5824
Igarashi K, Hiraga S, Yura T (1967) A deoxythymidine kinase deficient mutant of Escherichia coli. II. Mapping and transduction studies with phage 80. Genetics 57:643–654
Kaehler R, Strauss M, Kiessling U (1984) A well transformable E. coli tdk-strain-suitable for direct rescue of tk gene plasmids from mammalian cells. Biomed Biochim Acta 43:K25-K29
Kafatos FC, Jones Cw, Efstratiadis A (1979) Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucl Acids Res 7:1541–1551
Kiessling U, Platzer M, Strauss M (1984) Plasmid rescue—a tool for reproducible recovery of genes from transfected mammalian cells? Mol Gen Genet 193:513–519
Lacy E, Roberts S, Evan EP, Burtenshaw MD, Costantini FD (1983) A foreign β-globin gene in transgenic mice: Integration at abnormal chromosomal positions and expression in inappropriate tissues. Cell 34:343–358
Lund L, Grosveld FG, Flavell RA (1982) Isolation of transforming DNA by cosmid rescue. Proc Natl Acad Sci USA 79:520–524
Lusky M, Botchan MR (1984) Characterization of the bovine papilloma virus plasmid maintenance sequence. Cell 36:391–401
Maniatis T, Fritsch EF, Sambrook J (1982) A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor
McKnight GS, Hammer RE, Kuenzel EA, Brinster RL (1983) Expression of the chicken transferrin gene in trasgenic mice. Cell 34:335–341
Merrith GF, Hauschka SD, McKnight SL (1984) tk enzyme expression in differentiating muscle cells is regulated through an internal segment of the cellular tk gene. Mol Cell Biol 4:1777–1784
Palmiter RD, Chen HY, Brinster RL (1982) Differential regulation of metallothionein-thymidine kinase fusion genes in transgenic mice and their offspring. Cell 29:701–710
Palmiter RD, Wilkie TM, Chen HY, Brinster RL (1984) Transmission distortion and mosaicism in an unusual transgenic mouse pedigree. Cell 36:869–877
Perucho M, Hanahan D, Lipsich L, Wigler M (1980a) Isolation of the chicken thymidine kinase gene by plasmid rescue. Nature 285:207–210
Perucho M, Hanahan D, Wigler H (1980b) Genetic and physical linkage of exogenous sequences in transformed cells. Cell 22:309–317
Rigby PWJ, Brackman JK, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation. J Mol Biol 113:237–251
Spandidos DA, Paul J (1982) Transfer of human globin genes to erythroleukemic mouse cells. EMBO J 1:15–20
Stewart TA, Wagner EF, Mintz B (1982) Human β-globin gene sequences injected into mouse eggs, retained in adults, and transmitted to progeny. Science 217:1046–1048
Strauss M, Kiessling U (1983) Rescue of genes from mammalian cells by functional selection in E. coli. 15th FEBS Meeting, Brussels, Book of Abstracts, p 109
Strauss M, Kiessling U, Kaehler R (1983) Plasmids expressing the Herpes simplex virus thymidine kinase gene in mammalian and bacterial cells. Mol Gen Genet 191:154–157
Summers WC, Summers WP (1977) 125I Deoxycytidine used in a rapid, sensitive and specific assay for Herpes simplex virus type 1 thymidine kinase. J Virol 24:314–318
Wagner EF, Covarrubias L, Stewart TA, Mintz B (1984) Prenatal lethalities in mice homozygous for human growth hormone gene sequences integrated im the gern line. Cell 35:647–655
Wahl GM, Stern M, Stark GR (1979) Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci USA 76:3683–3687
Wigler M, Pellicer A, Silverstein S, Axel R (1978) Biochemical transfer of single-copy eukaryotic genes using total cellular DNA as donor. Cell 14:725–731
Wigler M, Silverstein S, Les LS, Pellicer A, Cheng YC, Axel R (1977) Transfer of purified Herpes virus thymidine kinase gene to cultured mouse cells. Cell 11:223–232
Yang RCA, Lis J, Wu R (1979) Elution of DNA from agarose gels after electrophoresis. Methods Enzymol 68:176–182
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Communicated by H. Böhme
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Kiessling, U., Becker, K., Strauss, M. et al. Rescue of a tk-plasmid from transgenic mice reveals its episomal transmission. Molec. Gen. Genet. 204, 328–333 (1986). https://doi.org/10.1007/BF00425518
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DOI: https://doi.org/10.1007/BF00425518