Vol. 27, 2009, Special Issue 2: S2-

Vol. 27, 2009, Special Issue 2: S2-75–S2-81 Czech J. Food Sci.





(bp)

500

200

162

100

Figure 7. Amplification of CaMV 35S promoter (162 bp). Line contents: M – HindIII DNA ladder (Fermentas, Canada); 1 – ≥ 0.03% positive control (BF410a); 2 – 0.1% positive control (BF410b); 3 – 1% positive control (BF410d); 4 – pa-paya flesh (isolation A); 5 – papaya flesh (isolation B); 6 – papaya stone (isolation A); 7 – papaya stone (isolation B); 8 – extraction control; 9 – MasterMix control




(bp)

500

200

162

100


Figure 8. Amplification of CaMV 35S promoter (162 bp). Line contents: M – HindIII DNA ladder (Fermentas, Can-ada); 1 – ≥ 0.03% positive control (BF410a); 2 – 0.1% po-sitive control (BF410b); 3 – 1% positive control (BF410d); 4 – candied papaya (isolation A); 5 – candied papaya (iso-lation B); 6 – extraction control; 7 – MasterMix control


result as an appropriate control, no 35S CaMV promoter sequence was detected in any assayed papaya material (Figures 7 and 8).


CONCLUSIONS

We verified and applied suitable protocol for DNA isolation from fresh papaya fruit and opti-mised the protocol for amplificable DNA isolation from candied fruits. The procedures can be suc-cessfully used for control purposes and monitoring of papaya import. No GM papaya fruits fresh or candied was found on the market.


References

Anonymous (2003): EC Regulation 1829/2003: on ge-netically modified (GM) food and feed. Available at http://europa.eu/eur-lex/pri/en/oj/dat/2003/l_268/ l_26820031018en00010023.pdf (accessed 7. 9. 2009).


Anklam E., Gadani F., Heinze P., Pijnenburg H., Van Den Eede G. (2002): Analytical methods for detection and determination of genetically modified organisms in agricultural crops and plant-derived food products. European Food Research and Technology, 214: 3–26.

AgBios GM Database: Available at www.agbios.com (accessed 7. 9. 2009).

Busch U., Pecoraro S., Posthoff K. (2004): Estendorfer-Rinner S First time detection of a genetically modified papaya in Europe – Official complaint of a non author-ized genetically modified organism within the EU Source. Deutsche Lebensmittel-Rundschau, 100: 377–380.

Bustin S.A. (2000): Asolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. Journal of Molecular Endocrinology, 25: 169–193.

Chapela M.J., Sotelo C.G., Pérez-Martín R.I., Par-do M.A., Pérez-Villareal B., Gilardi P., Riese J. (2007): Comparison of DNA extraction methods from muscle of canned tuna for species identification. Food Control, 18: 1211–1215.

Demeke T., Ratnayaka I., Phan A. (2009): Effects of DNA extraction and purification methods on real-time quantitative PCR analysis of Roundup Ready soybean. Journal of AOAC International, 92: 1136–1144.

EN ISO 21571:2002: Foodstuffs – Methods of analysis for the detection of genetically modified organisms and derived products – Nucleic acid extraction.

Fitch M.M., Manshardt R.M., Gonsalves D., Slightom J.L., Sanford J.C. (1990): Stable transfor-mation of papaya via microprojectile bombardment. Plant Cell Reports, 9: 189–194.

Fitch M.M., Manshardt R .M., Gonsalves D., Slightom J.L., Sanford J.C. (1992): Virus resistant papaya derived from tissues bombarded with the coat protein gene of papaya ringspot virus. Bio/Technology, 10: 1466–1472.