• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Dec 1, 1991; 88(23): 10558–10562.

Gene transfer with subsequent removal of the selection gene from the host genome.


A general method of gene transfer that does not leave behind a selectable marker in the host genome is described. A luciferase gene was introduced into the tobacco genome by using the hygromycin phosphotransferase gene (hpt) as a linked selectable marker. Flanked by recombination sites from the bacteriophage P1 Cre/lox recombination system, the hpt gene was subsequently excised from the plant genome by the Cre recombinase. The Cre-catalyzed excision event in the plant genome was precise and conservative--i.e., without loss or alteration of nucleotides in the recombinant site. After removal of the Cre-encoding locus by genetic segregation, plants were obtained that had incorporated only the desired transgene. Gene transfer without the incorporation of antibiotic-resistance markers in the host genome should ease public concerns over the field release of transgenic organisms expressing such traits. Moreover, it would obviate the need for different selectable markers in subsequent rounds of gene transfer into the same host.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Goodman RM, Hauptli H, Crossway A, Knauf VC. Gene transfer in crop improvement. Science. 1987 Apr 3;236(4797):48–54. [PubMed]
  • Weising K, Schell J, Kahl G. Foreign genes in plants: transfer, structure, expression, and applications. Annu Rev Genet. 1988;22:421–477. [PubMed]
  • Gasser CS, Fraley RT. Genetically engineering plants for crop improvement. Science. 1989 Jun 16;244(4910):1293–1299. [PubMed]
  • Perlak FJ, Deaton RW, Armstrong TA, Fuchs RL, Sims SR, Greenplate JT, Fischhoff DA. Insect resistant cotton plants. Biotechnology (N Y) 1990 Oct;8(10):939–943. [PubMed]
  • Abel PP, Nelson RS, De B, Hoffmann N, Rogers SG, Fraley RT, Beachy RN. Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science. 1986 May 9;232(4751):738–743. [PubMed]
  • Tumer NE, O'connell KM, Nelson RS, Sanders PR, Beachy RN, Fraley RT, Shah DM. Expression of alfalfa mosaic virus coat protein gene confers cross-protection in transgenic tobacco and tomato plants. EMBO J. 1987 May;6(5):1181–1188. [PMC free article] [PubMed]
  • Loesch-Fries LS, Merlo D, Zinnen T, Burhop L, Hill K, Krahn K, Jarvis N, Nelson S, Halk E. Expression of alfalfa mosaic virus RNA 4 in transgenic plants confers virus resistance. EMBO J. 1987 Jul;6(7):1845–1851. [PMC free article] [PubMed]
  • Hemenway C, Fang RX, Kaniewski WK, Chua NH, Tumer NE. Analysis of the mechanism of protection in transgenic plants expressing the potato virus X coat protein or its antisense RNA. EMBO J. 1988 May;7(5):1273–1280. [PMC free article] [PubMed]
  • van Dun CM, Bol JF. Transgenic tobacco plants accumulating tobacco rattle virus coat protein resist infection with tobacco rattle virus and pea early browning virus. Virology. 1988 Dec;167(2):649–652. [PubMed]
  • Lawson C, Kaniewski W, Haley L, Rozman R, Newell C, Sanders P, Tumer NE. Engineering resistance to mixed virus infection in a commercial potato cultivar: resistance to potato virus X and potato virus Y in transgenic Russet Burbank. Biotechnology (N Y) 1990 Feb;8(2):127–134. [PubMed]
  • Angenent GC, Van den Ouweland JM, Bol JF. Susceptibility to virus infection of transgenic tobacco plants expressing structural and nonstructural genes of tobacco rattle virus. Virology. 1990 Mar;175(1):191–198. [PubMed]
  • Golemboski DB, Lomonossoff GP, Zaitlin M. Plants transformed with a tobacco mosaic virus nonstructural gene sequence are resistant to the virus. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6311–6315. [PMC free article] [PubMed]
  • Craig NL. The mechanism of conservative site-specific recombination. Annu Rev Genet. 1988;22:77–105. [PubMed]
  • Dale EC, Ow DW. Intra- and intermolecular site-specific recombination in plant cells mediated by bacteriophage P1 recombinase. Gene. 1990 Jul 2;91(1):79–85. [PubMed]
  • Odell J, Caimi P, Sauer B, Russell S. Site-directed recombination in the genome of transgenic tobacco. Mol Gen Genet. 1990 Sep;223(3):369–378. [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]
  • Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. [PMC free article] [PubMed]
  • Kaster KR, Burgett SG, Rao RN, Ingolia TD. Analysis of a bacterial hygromycin B resistance gene by transcriptional and translational fusions and by DNA sequencing. Nucleic Acids Res. 1983 Oct 11;11(19):6895–6911. [PMC free article] [PubMed]
  • Ow DW, DE Wet JR, Helinski DR, Howell SH, Wood KV, Deluca M. Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science. 1986 Nov 14;234(4778):856–859. [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Hoess RH, Ziese M, Sternberg N. P1 site-specific recombination: nucleotide sequence of the recombining sites. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3398–3402. [PMC free article] [PubMed]
  • Sauer B. Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1987 Jun;7(6):2087–2096. [PMC free article] [PubMed]
  • Sauer B, Henderson N. Cre-stimulated recombination at loxP-containing DNA sequences placed into the mammalian genome. Nucleic Acids Res. 1989 Jan 11;17(1):147–161. [PMC free article] [PubMed]
  • Weatherall DJ. Gene therapy in perspective. Nature. 1991 Jan 24;349(6307):275–276. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...