Skip to main content
SpringerLink
Log in

Generation of transgenic plants expressing plasma membrane-bound antibodies to the environmental pollutant microcystin-LR

  • Brief Communication
  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

In this paper we describe the engineering and regeneration of transgenic tobacco plants expressing a recombinant plasma membrane-retained antibody specific to microcystin-LR (MC-LR), the environmental toxin pollutant produced by cyanobacteria. The antibody was created by a genetic fusion of the antigen binding regions of the microcystin-specific single chain antibody, 3A8, with the constant regions from the murine IgG1κ, Guy’s 13, including a membrane retention sequence at the C-terminal end of the antibody heavy chain. The antibody produced in the leaves was shown to be functional by binding to MC-LR in an ELISA with antibody yields in transgenic plant leaves reaching a maximum of 1.2 μg g−1 leaf f.wt (0.005% total soluble protein). Antibody-MC-LR complexes formed in leaves after addition of MC-LR to hydroponic medium around the roots of transgenic plant cultures.

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

Fig. 1
Fig. 2
Fig. 3

References

  • Drake PMW, Chargelegue D, Vine ND, van Dolleweerd CJ, Obregon P, Ma JKC (2002) Transgenic plants expressing antibodies: a model for phytoremediation. FASEB J 16:1855–1860

    Article  PubMed  CAS  Google Scholar 

  • Drake PMW, Barbi T, Drever MR, van Dolleweerd CJ, Porter AJR, Ma JKC (2010) Generation of transgenic plants expressing antibodies to the environmental pollutant microcystin-LR. FASEB J 24:882–890

    Article  PubMed  CAS  Google Scholar 

  • Hamvas M, Mathe C, Molnar E, Vasas G, Grigorszky I, Borbely G (2003) Microcystin-LR alters the growth, anthocyanin content and single-stranded DNase enzyme activities in Sinapis alba L. seedlings. Aquat Toxicol 62:1–9

    Article  Google Scholar 

  • Hanahan D (1983) Studies on transformation of Escherichia-Coli with plasmids. J Mol Biol 166:557–580

    Article  PubMed  CAS  Google Scholar 

  • Harada K, Oshikata M, Uchida H, Suzuki M, Kondo F, Sato K et al (1996) Detection and identification of microcystins in the drinking water of Haimen City, China. Nat Toxins 4:277–283

    Article  PubMed  CAS  Google Scholar 

  • Hassan S, van Dolleweerd CJ, Ioakeimidis F, Keshavarz-Moore E, Ma JKC (2008) Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants. Plant Biotechnol J 6:733–748

    Article  PubMed  CAS  Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general-method for transferring genes into plants. Science 227:1229–1231

    Article  CAS  Google Scholar 

  • Horton RM, Hunt HD, Ho SN, Pullen JK, Pease LR (1989) Engineering hybrid genes without the use of restriction enzymes—gene-splicing by overlap extension. Gene 77:61–68

    Article  PubMed  CAS  Google Scholar 

  • Kurki-Helasmo K, Meriluoto J (1998) Microcystin uptake inhibits growth and protein phosphatase activity in mustard (Sinapis alba L.) seedlings. Toxicon 36:1921–1926

    Article  PubMed  CAS  Google Scholar 

  • Ma JKC, Lehner T, Stabila P, Fux CI, Hiatt A (1994) Assembly of monoclonal-antibodies with Igg1 and Iga heavy-chain domains in transgenic tobacco plants. Eur J Immunol 24:131–138

    Article  PubMed  CAS  Google Scholar 

  • Mackintosh C, Beattie KA, Klumpp S, Cohen P, Codd GA (1990) Cyanobacterial Microcystin-Lr is a potent and specific inhibitor of Protein Phosphatase-1 and Phosphatase-2A from both mammals and higher-plants. FEBS Lett 264:187–192

    Article  PubMed  CAS  Google Scholar 

  • McElhiney J, Lawton LA, Leifert C (2001) Investigations into the inhibitory effects of microcystins on plant growth, and the toxicity of plant tissues following exposure. Toxicon 39:1411–1420

    Article  PubMed  CAS  Google Scholar 

  • McElhiney J, Drever M, Lawton LA, Porter AJ (2002) Rapid isolation of a single-chain antibody against the cyanobacterial toxin microcystin-LR by phage display and its use in the immunoaffinity concentration of microcystins from water. Appl Environ Microb 68:5288–5295

    Article  CAS  Google Scholar 

  • Meagher RB, Heaton ACP (2005) Strategies for the engineered phytoremediation of toxic element pollution: mercury and arsenic. J Ind Microbiol Biotechnol 32:502–513

    Article  PubMed  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth bioassays with tobacco tissue cultures. Plant Physiol 15:473–497

    Article  CAS  Google Scholar 

  • Rogers SG, Klee HJ, Horsch RB, Fraley RT (1987) Improved vectors for plant transformation—expression cassette vectors and new selectable markers. Methods Enzymol 153:253–277

    Article  CAS  Google Scholar 

  • Staub JM, Garcia B, Graves J, Hajdukiewicz PTJ, Hunter P, Nehra N et al (2000) High-yield production of a human therapeutic protein in tobacco chloroplasts. Nat Biotechnol 18:333–338

    Article  PubMed  CAS  Google Scholar 

  • Ueno Y, Nagata S, Tsutsumi T, Hasegawa A, Watanabe MF, Park HD et al (1996) Detection of microcystins, a blue-green algal hepatotoxin, in drinking water sampled in Haimen and Fusui, endemic areas of primary liver cancer in China, by highly sensitive immunoassay. Carcinogenesis 17:1317–1321

    Article  PubMed  CAS  Google Scholar 

  • Vine ND, Drake P, Hiatt A, Ma JKC (2001) Assembly and plasma membrane targeting of recombinant immunoglobulin chains in plants with a murine immunoglobulin transmembrane sequence. Plant Mol Biol 45:159–167

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank to Sir Joseph Hotung for funding PMWD and JK-CM. TB was funded by a joint St George’s University of London/Royal Holloway University of London PhD studentship.

Author information

Authors and Affiliations

  1. Molecular Immunology Unit, Centre for Infection, Division of Clinical Sciences, St. George’s University of London, Cranmer Terrace, room 2.148 Floor 2 Jenner Wing, London, SW17 0RE, UK

    Tommaso Barbi, Pascal M. W. Drake, Craig J. van Dolleweerd & Julian K-C. Ma

  2. Department of Molecular and Cell Biology, Foresterhill School of Medical Sciences, University of Aberdeen, Aberdeen, UK

    Matthew Drever & Andrew R. Porter

Authors
  1. Tommaso Barbi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pascal M. W. Drake
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Matthew Drever
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Craig J. van Dolleweerd
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Andrew R. Porter
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Julian K-C. Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pascal M. W. Drake.

Additional information

Tommaso Barbi, Pascal M.W. Drake these authors contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Barbi, T., Drake, P.M.W., Drever, M. et al. Generation of transgenic plants expressing plasma membrane-bound antibodies to the environmental pollutant microcystin-LR. Transgenic Res 20, 701–707 (2011). https://doi.org/10.1007/s11248-010-9431-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11248-010-9431-4

Keywords

Search

Navigation