Skip to main content
Log in

Characterization of polyadenylated cryIA(b) transcripts in maize MON810 commercial varieties

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

The Zea mays L. event MON810 is one of the major commercialized genetically modified crops. The inserted expression cassette has a 3′ truncation partially affecting the cryIA(b) coding sequence, resulting in the lack of the NOS terminator, with transcription of the transgene reported to read-through 3′-past the truncation site. Here, we demonstrate that the cryIA(b) transgene gives rise to a variety of polyadenylated transcripts of different sizes that extend to around 1 kbp downstream the truncation site. A Stop codon at position +7 downstream the truncation site indicates the production of a transgenic protein with two additional amino acids; which is compatible with the reported size of the CryIA(b) protein in MON810. There is no evidence of the existence of other translated products. Several main 3′ transcription termination regions were detected close to the truncation site and in the transgene 3′ flanking sequence. Next to these main termination sites, we identified some sequence motifs that could potentially act as 3′-end-processing elements and drive termination of the transgene transcripts. The MON810 transgene has been introduced into different commercial varieties through breeding programs. Here, we demonstrate that there are no significant differences among the levels of transgene mRNA accumulation, major transcript sizes and 3′ termini profiles comparing a number of MON810 commercial varieties grown under similar environmental conditions. Commercial varieties of this event appear to be stable in terms of transgene expression.

Similar cryIA(b) RACE 3’-PCR profiles were obtained with mRNA from six MON810 commercial varieties: superposition of electropherograms and schematic representation of the length of transgene mRNA molecules.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

CaMV:

Cauliflower mosaic virus

BAC:

Bacterial artificial chromosome

NOS:

Nopaline synthetase

GMO:

Genetically modified organism

mRNA:

Messenger ribonucleic acid

CRM:

Certified reference material

IRMM:

Institute for Reference Materials and Measurements

Real-time RT-PCR:

Reverse transcription coupled to real-time PCR

PCR:

Polymerase chain reaction

cDNA:

Complementary deoxyribonucleic acid

RACE 3′-PCR:

Rapid amplification of cDNA ends

References

  1. Hernández M, Pla M, Esteve T, Prat S, Puigdomènech P, Ferrando A (2003) Transgenic Res 12(2):179–189

    Article  Google Scholar 

  2. Rosati A, Bogani P, Santarlasci A, Buiatti M (2008) Plant Mol Biol 67:271–281

    Article  CAS  Google Scholar 

  3. Rasmussen R (2001) In: Meuer S, Wittwer C, Nakagawara K (eds) Rapid Cycle real-time PCR, methods and applications. Springer, Berlin,G

    Google Scholar 

  4. Arumuganathan K, Earle ED (1991) Plant Mol Biol Rep 9:208–218

    Article  CAS  Google Scholar 

  5. Sambrook J, MacCallum P, Russell D (2000) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press

  6. Bubner B, Gase K, Baldwin IT (2004) BMC Biotechnol 4:14–20

    Article  Google Scholar 

  7. Rothnie HM (1996) 32(1-2):43-61

  8. Li Q, Hunt G (1997) Plant Physiol 115:321–325

    Article  CAS  Google Scholar 

  9. Kohli A, Twyman R, Abranches R, Wegel E, Stoger E, Christou P (2003) Plant Mol Biol 52:247–258

    Article  CAS  Google Scholar 

  10. Zhang J, Cai L, Cheng J, Mao H, Fan X, Meng Z, Chan KM, Zhang H, Qi J, Ji L, Hong Y (2008) Transgenic Res 17(2):293–306

    Article  CAS  Google Scholar 

  11. Windels P, Taverniers I, Depicker A, Van Bockstaele E, De Loose M (2001) Eur Food Res Technol 213:107–111

    Article  CAS  Google Scholar 

  12. Rang A, Linke B, Jansen B (2005) Eur Food Res Technol 220:438–443

    Article  CAS  Google Scholar 

  13. Kashkush K, Feldman M, Levy AA (2003) Nat Genet 33:102–106

    Article  CAS  Google Scholar 

  14. Kashkush K, Khasdan V (2007) Genetics 177:1975–1985

    Article  CAS  Google Scholar 

  15. Hernández-Pinzón I, de Jesús E, Santiago N, Casacuberta JM (2009) J Mol Evol in press. doi:10.1007/s00239-009-9204-y

  16. Vitte C, Bennetzen JL (2006) Proc Natl Acad Sci USA 103(47):17638–17643

    Article  CAS  Google Scholar 

  17. Hellmann H, Estelle M (2002) Science 297(5582):793–797

    Article  CAS  Google Scholar 

  18. Kee Y, Huibregtse JM (2007) Biochem Biophys Res Commun 354(2):329–333

    Article  CAS  Google Scholar 

  19. Downes BP, Stupar RM, Gingerich DJ, Vierstra RD (2003) The Plant Journal 35(6):729–742

    Article  CAS  Google Scholar 

  20. Hunt AG (2008) Curr Top Microbiol Immunol 326:151–177

    Article  CAS  Google Scholar 

  21. Edmonds M (2002) Prog Nucleic Acid Res Mol Biol 71:285–389

    Article  CAS  Google Scholar 

  22. Loke JC, Stahlberg EA, Strenski DG, Haas BJ, Wood PC, Li QQ (2005) Plant Physiol 138(3):1457–1468

    Article  CAS  Google Scholar 

  23. Dong H, Deng Y, Chen J, Wang S, Peng S, Dai C, Fang Y, Shao J, Lou Y, Li D (2007) Gene 389:107–113

    Article  Google Scholar 

  24. Ji G, Zheng J, Shen Y, Wu X, Jiang R, Lin Y, Loke J, Davis K, Reese G, Li Q (2007) BMC Bioinformatics 8:43

    Article  Google Scholar 

  25. Shen Y, Ji G, Haas BJ, Wu X, Zheng J, Reese GJ, Li QQ (2008) Nucleic Acids Res 36(9):3450–3161

    Article  Google Scholar 

  26. Nguyen HT, Jehle JA (2007) J Plant Dis Protect 114:820–887

    Google Scholar 

  27. Coll A, Nadal A, Palaudelmàs M, Messeguer J, Melé E, Puigdomènech P, Pla M (2008) Plant Mol Biol 68:105–117

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was financially supported by the EC FP6 project “Co-Extra” and by the Spanish MEC project with ref. AGL2007-65903/AGR.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Maria Pla.

Rights and permissions

Reprints and permissions

About this article

Cite this article

La Paz, J.L., Vicient, C., Puigdomènech, P. et al. Characterization of polyadenylated cryIA(b) transcripts in maize MON810 commercial varieties. Anal Bioanal Chem 396, 2125–2133 (2010). https://doi.org/10.1007/s00216-009-3176-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-009-3176-z

Keywords

Navigation