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.

Hasil (Bahasa Indonesia) 1:

Vol. 27, 2009, khusus edisi 2: S2-75-S2-81 Ceko J. makanan Sci.(bp)500200162100Gambar 7. Amplifikasi promotor CaMV 35S (162 bp). Garis isi: M-HindIII DNA tangga (Fermentas, Kanada); 1 – ≥ 0.03% positif kontrol (BF410a); 2-0.1% positif kontrol (BF410b); 3-1% kontrol positif (BF410d); 4-pa-paya daging (isolasi A); 5 – pepaya daging (isolasi B); 6-pepaya batu (isolasi A); 7-pepaya batu (isolasi B); 8-ekstraksi kontrol; 9 – MasterMix kontrol (bp)500200162100Gambar 8. Amplifikasi promotor CaMV 35S (162 bp). Garis isi: M-HindIII DNA tangga (Fermentas, Can-ada); 1 – ≥ 0.03% positif kontrol (BF410a); 2-0.1% kontrol po-sitive (BF410b); 3-1% kontrol positif (BF410d); 4-manisan pepaya (isolasi A); 5 – manisan pepaya (iso-hal yang sanggat menarik B); 6 – ekstraksi kontrol; 7-MasterMix kontrolhasil sebagai kontrol yang sesuai, tidak ada CaMV 35S promotor urutan dideteksi pada bahan diuji pepaya (angka 7 dan 8).KESIMPULANKami diverifikasi dan diterapkan protokol cocok untuk isolasi DNA dari buah-buahan segar pepaya dan opti-mised protokol untuk isolasi DNA amplificable dari manisan buah-buahan. Prosedur dapat suc-cessfully digunakan untuk tujuan pengendalian dan pemantauan pepaya impor. Tidak ada GM buah pepaya segar atau manisan ditemukan di pasar.ReferensiAnonim (2003): EC peraturan 1829/2003: ge netically diubah (GM) makanan dan pakan. Tersedia di http://europa.eu/eur-lex/pri/en/oj/dat/2003/l_268/ l_26820031018en00010023.pdf (diakses 7. 9. 2009). Anklam E., Gadani F., Heinze P., Pijnenburg H., Van Den Eede G. (2002): Metode analisis untuk Deteksi dan penentuan modified organisme dalam tanaman pertanian dan produk makanan tanaman-berasal. Makanan Eropa riset dan teknologi, 214: 3 – 26.AgBios GM Database: Tersedia di www.agbios.com (diakses 7. 9. 2009).Busch U., Pecoraro S., K. Posthoff (2004): Estendorfer-Rinner pertama S waktu deteksi pepaya transgenik di Eropa-pengaduan resmi dari penulis bebas-ized modified organisme dalam sumber Uni Eropa. Deutsche Lebensmittel-Rundschau, 100: 377-380.Bustin S.A. (2000): Asolute kuantifikasi mRNA menggunakan tes reaksi berantai polimerase real-time transkripsi terbalik. Jurnal endokrinologi molekuler, 25: 169-193.Chapela M.J. Sotelo perkataan, Pérez-Martín R.I., Par-do M.A., B. Pérez-Villareal, Gilardi P., Riese J. (2007): Metode ekstraksi perbandingan DNA dari otot kaleng tuna untuk identifikasi spesies. Kontrol makanan, 18: 1211-1215.T. Demeke, Ratnayaka I., Phan A. (2009): Efek dari DNA ekstraksi dan pemurnian metode analisis PCR kuantitatif real-time Roundup siap kedelai. Jurnal AOAC International, 92: 1136-1144.EN ISO 21571:2002: bahan makanan – metode analisis untuk deteksi organisme yang dimodifikasi secara genetik dan produk turunan – asam nukleat ekstraksi.Fitch M.M., Manshardt R.M., Gonsalves D., Jl Slightom, Sanford J.C. (1990): Stabil transfor-Masi pepaya melalui microprojectile pemboman. Tanaman sel laporan, 9: 189-194.Fitch M.M., Manshardt R.M., D. Gonsalves, Jl Slightom, Sanford J.C. (1992): Tahan Virus pepaya berasal dari jaringan dibombardir dengan mantel protein gen pepaya ringspot virus. Bioteknologi, 10: 1466-1472.