Skip to main content
SpringerLink
Log in

Agroinfiltration as a Tool for Transient Expression of cre Recombinase in vivo

  • Short communication
  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

Agroinfiltration was used to express transiently cre recombinase from bacteriophage P1 in planta. Activation of gfp expression after cre-mediated excision of a bar intervening sequence served as a marker to monitor site-specific recombination events in lox-target N. benthamiana plants. Gfp expressing regenerants from A. tumefaciens infiltrated leaves were obtained with an efficiency of about 34%. In 20% of the regenerants bar gene excision was due to the expression of stably integrated cre gene, whereas in 14% of plants site-specific recombination was a consequence of transient cre expression. Phenotypic and molecular data indicated that the recombined state has been transferred to the T1 generation. These results demonstrate the suitability of agroinfiltration for the expression of cre recombinase in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • H Araki A Jearnpipatkul H Tatsumi T Sakurai K Ushio T Muta Y Oshima (1985) ArticleTitleMolecular and functional organisation of the yeast plasmid pSR1 J Mol Biol 182 191–203 Occurrence Handle10.1016/0022-2836(85)90338-9 Occurrence Handle3889347

    Article  PubMed  Google Scholar 

  • CC Bayley M Morgan EC Dale DW Ow (1992) ArticleTitleExchange of gene activity in transgenic plants catalyzed by the Cre-lox site-specific recombination system Plant Mol Biol 18 353–361 Occurrence Handle10.1007/BF00034962 Occurrence Handle1310059

    Article  PubMed  Google Scholar 

  • JR Broach VR Guarascio M Jayaram (1982) ArticleTitleRecombination within the yeast 2-μm circle is site-specific Cell 29 227–234 Occurrence Handle10.1016/0092-8674(82)90107-6 Occurrence Handle6286142

    Article  PubMed  Google Scholar 

  • ER Coppoolse MJ Vroomen Particlede D Roelofs J Smit F Gennip Particlevan BJM Hersmus et al. (2003) ArticleTitleCre recombinase expression can result in phenotypic aberrations in plants Plant Mol Biol 51 263–279 Occurrence Handle10.1023/A:1021174726070 Occurrence Handle12602884

    Article  PubMed  Google Scholar 

  • S Corneille K Lutz Z Svab P Maliga (2001) ArticleTitleEfficient elimination of selectable marker genes from the plastid genome by the Cre-lox site-specific recombination system Plant J 27 171–178 Occurrence Handle10.1046/j.1365-313x.2001.01068.x Occurrence Handle11489194

    Article  PubMed  Google Scholar 

  • EC Dale DW Ow (1990) ArticleTitleIntra and intermolecular site-specific recombination in plant cells mediated by bacteriophage P1 recombinase Gene 91 79–85 Occurrence Handle10.1016/0378-1119(90)90165-N Occurrence Handle2205542

    Article  PubMed  Google Scholar 

  • EC Dale DW Ow (1991) ArticleTitleGene transfer with subsequent removal of the selection gene from the host genome Proc Natl Acad Sci USA 88 10558–10562 Occurrence Handle1660141

    PubMed  Google Scholar 

  • CD Day E Lee J Kobayashi LD Holappa H Albert DW Ow (2000) ArticleTitleTransgene integration into the same chromosome location can produce alleles that express at a predictable level, or alleles that are differentially silenced Gene Dev 14 2869–2880 Occurrence Handle10.1101/gad.849600 Occurrence Handle11090134

    Article  PubMed  Google Scholar 

  • SV Gelvin (2003) ArticleTitleAgrobacterium-mediated plant transformation: the biology behind the “gene-jockeying” tool Microbiol. Mol Biol Rev 67 16–37 Occurrence Handle10.1128/MMBR.67.1.16-37.2003 Occurrence Handle12626681

    Article  PubMed  Google Scholar 

  • AP Gleave DS Mitra SR Mudge B.A.M Morris (1999) ArticleTitleSelectable marker-free transgenic plants without sexual crossing: transient expression of cre recombinase and use of a conditional lethal dominant gene Plant Mol Biol 40 223–235 Occurrence Handle10.1023/A:1006184221051 Occurrence Handle10412902

    Article  PubMed  Google Scholar 

  • TTC Hoa BB Bong E Huq TK Hodges (2002) ArticleTitleCre-lox site-specific recombination controls the excision of a transgene from the rice genome Theor Appl Genet 104 518–525 Occurrence Handle10.1007/s001220100748 Occurrence Handle12582653

    Article  PubMed  Google Scholar 

  • RH Hoess K Abremski (1985) ArticleTitleMechanism of strand cleavage and exchange in the Cre-lox site-specific recombination system J Mol Biol 181 351–362 Occurrence Handle10.1016/0022-2836(85)90224-4 Occurrence Handle3856690

    Article  PubMed  Google Scholar 

  • B-J Janssen RC Gardner (1989) ArticleTitleLocalised transient expression of gusA in leaf discs following cocultivation with Agrobacterium Plant Mol Biol 14 61–72 Occurrence Handle10.1007/BF00015655

    Article  Google Scholar 

  • J Kapila R Rycke Particlede M Montagu Particlevan G Angenon (1997) ArticleTitleAn Agrobacterium- mediated transient gene expression system for intact leaves Plant Sci 122 101–108 Occurrence Handle10.1016/S0168-9452(96)04541-4

    Article  Google Scholar 

  • L Kopertekh G Jüttner J Schiemann (2004a) ArticleTitleSite-specific recombination induced in transgenic plants by PVX virus vector expressing bacteriophage P1 recombinase Plant Science 166 485–492 Occurrence Handle10.1016/j.plantsci.2003.10.018

    Article  Google Scholar 

  • L Kopertekh G Jüttner J Schiemann (2004b) ArticleTitlePVX-Cre-mediated marker gene elimination from transgenic plants Plant Mol Biol 55 491–500 Occurrence Handle10.1007/s11103-004-0237-8

    Article  Google Scholar 

  • HA Koshinsky E Lee DW Ow (2000) ArticleTitleCre-lox site-specific recombination between Arabidopsis and tobacco chromosomes Plant J 23 715–222 Occurrence Handle10.1046/j.1365-313x.2000.00839.x Occurrence Handle10998183

    Article  PubMed  Google Scholar 

  • ZZ Liu JL Wang X Huang WH Xu ZM Liu RX Fang (2003) ArticleTitleThe promoter of a rice glycine-rich protein gene, Osgrp-2, confers vascular-specific expression in transgenic plants Planta 216 824–833 Occurrence Handle12624770

    PubMed  Google Scholar 

  • LA Lyznik WJ Gordon-Kamm Y Tao (2003) ArticleTitleSite-specific recombination for genetic engineering in plants Plant Cell Rep 21 925–932 Occurrence Handle10.1007/s00299-003-0616-7 Occurrence Handle12835900

    Article  PubMed  Google Scholar 

  • LA Lyznik L Hirayama KV Rao A Abad TK Hodges (1995) ArticleTitleHeat-inducible expression of FLP gene in maize cells Plant J 8 177–186 Occurrence Handle10.1046/j.1365-313X.1995.08020177.x Occurrence Handle7670503

    Article  PubMed  Google Scholar 

  • LA Lyznik KV Rao TK Hodges (1996) ArticleTitleFLP-mediated recombination of FRT sites in the maize genome Nucl Acids Res 24 3784–3789 Occurrence Handle10.1093/nar/24.19.3784 Occurrence Handle8871559

    Article  PubMed  Google Scholar 

  • S Maeser R Kahmann (1991) ArticleTitleThe Gin recombinase of phage Mu can catalyse site-specific recombination in plant protoplasts Mol Gen Genet 230 170–176 Occurrence Handle10.1007/BF00290665 Occurrence Handle1836050

    Article  PubMed  Google Scholar 

  • J Odell P Caimi B Sauer S Russell (1990) ArticleTitleSite-directed recombination in the genome of transgenic tobacco Mol Gen Genet 223 369–378 Occurrence Handle10.1007/BF00264442 Occurrence Handle2176714

    Article  PubMed  Google Scholar 

  • DW Ow (2002) ArticleTitleRecombinase-directed plant transformation for the post-genomic era Plant Mol Biol 48 183–200 Occurrence Handle10.1023/A:1013718106742 Occurrence Handle11860209

    Article  PubMed  Google Scholar 

  • K Palanichelvam AB Cole M Shabababi JE Schoelz (2000) ArticleTitleAgroinfiltration of Cauliflower mosaic virus gene VI elicits hypersensitive response in Nicotiana species Mol Plant Microbe Interact 13 1275–1279 Occurrence Handle11059496

    PubMed  Google Scholar 

  • SR Scofield CM Tobias JP Rathjen JH Chang DT Lavelle RM Michelmore et al. (1996) ArticleTitleMolecular basis of gene-for-gene specificity in bacterial speck disease of tomato Science 274 2063–2065 Occurrence Handle10.1126/science.274.5295.2063 Occurrence Handle8953034

    Article  PubMed  Google Scholar 

  • V Srivastava DW Ow (2001) ArticleTitleSingle-copy primary transformants of maize obtained through the co-introduction of a recombinase-expressing construct Plant Mol Biol 46 561–566 Occurrence Handle10.1023/A:1010646100261 Occurrence Handle11516149

    Article  PubMed  Google Scholar 

  • K Sugita T Kasahara E Matsunaga H Ebinuma (2001) ArticleTitleA transformation vector for the production of marker-free transgenic plants containing a single copy transgene at high frequency Plant J 22 461–469 Occurrence Handle10.1046/j.1365-313X.2000.00745.x

    Article  Google Scholar 

  • C Vaquero M Sack J Chandler J Drossard F Schuster M Monecke et al. (1999) ArticleTitleTransient expression of a tumor-specific single-chain fragment and a chimeric antibody in tobacco leaves Proc Natl Acad Sci USA 96 11128–11133 Occurrence Handle10.1073/pnas.96.20.11128 Occurrence Handle10500141

    Article  PubMed  Google Scholar 

  • AC Vergunst LET Jansen PJJ Hooykaas (1998) ArticleTitleSite-specific integration of Agrobacterium T-DNA in Arabidopsis thaliana mediated by Cre recombinase Nucl Acids Res 26 2729–2734 Occurrence Handle10.1093/nar/26.11.2729 Occurrence Handle9592161

    Article  PubMed  Google Scholar 

  • AC Vergunst B Schrammeijer A Dulk-Ras Particleden CMT Vlaam Particlede TJG Regensburg-Tuink PJJ Hooykaas (2000) ArticleTitleVir B/D4 – dependent protein translocation from Agrobacterium into plant cells Science 290 979–982 Occurrence Handle10.1126/science.290.5493.979 Occurrence Handle11062129

    Article  PubMed  Google Scholar 

  • O Voinnet S Rivas P Mestre D Baulcombe (2003) ArticleTitleAn enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus Plant J 33 949–956 Occurrence Handle10.1046/j.1365-313X.2003.01676.x Occurrence Handle12609035

    Article  PubMed  Google Scholar 

  • W Zhang S Subbaro P Addae A Shen C Armstrong V Peschke et al. (2003) ArticleTitleCre/lox-mediated marker gene excision in transgenic maize (Zea mays L.) plants Theor Appl Genet 107 1157–1168 Occurrence Handle10.1007/s00122-003-1368-z Occurrence Handle14513214

    Article  PubMed  Google Scholar 

  • J Zuo Q-W Niu SG Moller N-H Chua (2001) ArticleTitleChemical-regulated, site-specific DNA excision in transgenic plants Nat Biotechnol 19 157–161 Occurrence Handle10.1038/84428 Occurrence Handle11175731

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Federal Biological Research Centre for Agriculture and Forestry, Institute for Plant Virology, Microbiology and Biosafety, Messeweg 11-12, D-38104, Braunschweig, Germany

    Lilya Kopertekh & Joachim Schiemann

Authors
  1. Lilya Kopertekh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joachim Schiemann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Joachim Schiemann.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kopertekh, L., Schiemann, J. Agroinfiltration as a Tool for Transient Expression of cre Recombinase in vivo. Transgenic Res 14, 793–798 (2005). https://doi.org/10.1007/s11248-005-8293-7

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11248-005-8293-7

Keywords

Search

Navigation