Identifikasi Gen Transgenik pada Produk Susu Bubuk Kedelai dan Susu Formula Soya dengan Metode PCR (Polymerase Chain Reaction)
Agustin Krisna Wardani(1*), Annisa Alirsyah(2), Ana Fauziah(3)
(1) Jurusan Teknologi Hasil Pertanian, Fakultas Teknologi Pertanian, Universitas Brawijaya, Jl. Veteran, Malang 65145
(2) Jurusan Teknologi Hasil Pertanian, Fakultas Teknologi Pertanian, Universitas Brawijaya, Jl. Veteran, Malang 65145
(3) Jurusan Teknologi Hasil Pertanian, Fakultas Teknologi Pertanian, Universitas Brawijaya, Jl. Veteran, Malang 65145
(*) Corresponding Author
Abstract
The need of soybean that reached up to 2,2 million tons per year has made Indonesia imports this commodity 1,62 million tons from countries that adopt Genetically Modified (GM) soybean. By the presence of GM soybeans in Indonesia, Genetically Modified Product (GMP) labelling has need to be done. Detection of GMP can be done by using PCR. The aim of this study was to determine the presence of trangenic genetic material in soy milk powder and soy formula products to classify as GMP. Another goal was to determine the optimum annealing temperature of the primers used. Based on this study, the optimum annealing temperature of the CaMV 35S primer and the EPSPS-CP4 primer was 60oC and 59oC. The NOS terminator primer’s optimum annealing temperature was not found. 6 soy milk powder samples and 5 soy formula samples are might be determined to be using transgenic soybeans due to the presence of EPSPS-CP4 genes and CaMV 35S promotor genes. Therefore, those 11 samples were classified as GMP.
ABSTRAK
Kebutuhan kedelai yang mencapai 2,2 juta ton/tahun memaksa Indonesia mengimpor sebanyak 1,62 juta ton. Sebagian besar kedelai impor berupa kedelai transgenik. Dengan munculnya kedelai transgenik di Indonesia, perlu adanya pelabelan Produk Rekayasa Genetika (PRG) untuk memenuhi hak-hak konsumen. Teknik yang dilakukan untuk mendeteksi PRG salah satunya menggunakan metode PCR. Penelitian ini bertujuan untuk mengetahui ada tidaknya gen transgenik pada produk susu bubuk kedelai dan formula soya, sehingga produk dapat digolongkan sebagai PRG atau tidak. Selain itu juga bertujuan untuk mengetahui suhu annealing optimum pada primer yang digunakan. Hasil penelitian didapatkan suhu annealing optimum primer CaMV 35S promotor adalah 60oC. Sedangkan untuk primer gen EPSPS-CP4 suhu annealing optimumnya 59oC. Untuk primer NOS terminator suhu annealing optimum tidak ditemukan. Dari amplifikasi DNA sampel, 6 sampel susu bubuk kedelai dan 5 sampel formula soya terdapat sisipan gen EPSPS-CP4 dan gen Promotor CaMV 35S. Dengan demikian 11 sampel tersebut dapat dikatakan sebagai PRG.
Kata kunci: Produk rekayasa genetika; PCR; formula soya; susu kedelai bubuk; kedelai transgenik
Keywords
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ACNFP. (2002). Monsanto Roundup Ready soybeans line 40-3-2. http://acnfp.food-.gov.uk. [15 Oktober 2015].
Bauer, T., Weller, P., Hammes, W.P. dan Hertel, C. (2003). The effect of processing on DNA degradation in food. Europan Food research and Technology 217: 338–343.
Carson, S. (2006). Manipulation and Expression of Recombinant DNA A Laboratory Manual Second Edition. Elsevier Academic Press, USA.
Chen, K., Han, H.dan Wang, X. (2012). A practicable detection system for genetically modified rice by SERS-barcoded nanosensors. Biosensors and Bioelectronics 34: 118–124.
Grunewald, H. (2003). Optimization of Polymerase Chain Reaction. Dalam: Bartlett, J.M.S. dan Stirling, D. (ed.). Methods in Molecular Biology Vol 226: PCR Protocols Second Edition. Humana Press, New Jersey.
Hammond B.G. dan Jez, J.M. (2011). Impact of food processing on the safety assessment for proteins introduced into biotechnology-derived soybean and corn crops. Food and Chemical Toxicology 49(4): 711–721.
Handoyo, D. dan Ari, R. (2000). Prinsip umum dan pelaksanaan polymerase chain reaction (PCR). Jurnal Unesa 9(1): 17–29.
Hardi, Y. (2009). Reaksi PCR. http://sciencebiotech.net/mengenal-pcr-polimerase-chain-reaction/. [16 Juni 2015].
Hadiarto, T., Utami, T.I.R., Pardal, S.J. dan Herman, M. (2001). Analisis molekuler gen pinii pada tanaman kedelai transgenik R2. Prosiding Seminar Hasil Penelitian Rintisan dan Bioteknologi Tanaman Bogor 133–140.
Ilhak, O.I. dan Arslan, A. (2007). Identification of meat species by polymerase chain reaction (PCR) technique. Turkish Journal Veterinary Animal 31(3): 159–163.
Folloni, S., Bellocchi, G., Kagkli, D.M., Benito, S.P., Aguilera, M., Mazzeo, A., Querci, M., Eede, G.V. dan Ermolli, M. (2011). Development of an ELISA reverse-based assay to assess the oresence of mycotoxin in cereal flour. Food Analytical Methods 4: 221–227.
Jankiewicz, A., Broll, H. dan Zagon, J. (1999). The official method for the detection of genetically modified soybeans: a semi-quantitative study of sensitivity limits with glyphosate-tolerant soybeans (Roundup Ready) and insect-resistant Bt maize (Mazimizer). European Food Research and Technology 209: 77–82.
Kilamanca, C.M. (2008). Sikap Konsumen Pasar Swalayan terhadap Produk Susu Kedelai di Kota Surakarta. Skripsi. Universitas Sebelas Maret, Surakarta.
Lipp, M., Bluth, A., Eyquem, F., Kruse, L., Schimmel, H., Eede, G.V. dan Anklam, E. (2001). Validation of a method based on polymerase chain reaction for the detection of genetically modified organisms in various processed foodstuffs. European Food Research and Technology 212(4): 497–504.
Meric, S. Cakir, O. Ari, S. dan Turqut, K.N. (2014). Detection of genetically modified maize and soybean in feed samples. Genetics and Molecular Research 13(1): 1160–1168.
Muladno (2010). Teknologi Rekayasa Genetika Edisi Kedua. Institut Pertanian Bogor Press, Bogor.
Nazir, M. (2005). Metode Penelitian. Ghalia Indonesia, Bogor.
Nguyen, T.C.T., Son, R., Raha, A.R., Lai, O.M. dan Michael C. (2009). Comparison of DNA extraction eficiencies using various methodes for the detection of genetically modified organisms (GMO). Journal International Food Research 16: 21–30.
Olson, P.S. (2015). Peran Tanaman Transgenik Semakin Nyata. http://tabloidsinartani.com/-content/read/peran-tanaman-transgenik-semakin-nyata/. [22 Agustus 2015].
Peccia, J. dan Hernadez, M. (2006). Incorporating polymerase chain reaction-based identification population characterization, and quantification of microorganisms into aerosol: A review. Atmospheric Environment 40(21): 3941–3961.
Pena, L. (2005). Trasngenic Plants: Methode and Protocol Methode in Molecular Biology Vol 286. Humana Press, New Jersey.
Putra, S.E.D., Tsuprykov, O., Websky, K.V., Ritter, T., Reichetzeder, C. dan Hocher, B. (2014). Dealing with large sample sizes: comparison of a new one spot dot blot method to Western blot. Clinical Laboratory 60: 1871–1877.
Rahmawati (2011). Identifikasi Gen Transgenik pada Kedelai Impor dan Tempe di Kota Malang.Skripsi. Universitas Brawijaya, Malang.
Randhawa, G.J. dan Firke, P.K. (2006). Detection of transgenes in genetically modified soybean and maize using polymerase chain reaction. Indian Journal of Biotechnology 5: 510–513.
Sambrook, J.S. dan Russel, D.W. (2001). Molecular Cloning: A Laboratory Manual. Volume 1-3,Third Edition. Cold Spring arbour Laboratory Press, New York.
Sari, E.P.K. dan Wardani, A.K. (2015). Deteksi molekuler cemaran daging babi pada bakso sapi di pasar tradisional kota malang menggunakan PCR (polymerase chain reaction). Jurnal Pangan dan Agroindustri 3(4): 1294–1301.
Satria (2015). Produksi Kedelai Nasional Rendah. http://ugm.ac.id/. [15 Oktober 2015].
Sawazaki, H.E., Duarte, A.P., Fuzatto, M.G., Sawazaki, E., Grandi, S.H.R., Ponte, J.F. dan Noguire, L. (2015). Identification and quantification of corn, soybean and cotton genetically modified by real time PCR. American Journal of Molecular Biology 5: 84–93.
Sorina, M. Maria, B.O., Carasela, P. dan Elena, B. (2006). The GM (genetically modified) detection and quantification in different corn samples. Journal of Horticulture Forestry and Biotechnology: 398–401.
Termos Fisher Scientific (2008). Nanodrop 1000 Spectrophotometer V.3.7 User Manual. www.nanodrop.com/library/nd-1000-v3.7-users-manual-8.5x11.pdf. [23 Oktober 2015].
Tung, N.C.T., Son, R., Raha, A.R., Lai, O.M. dan Clemente M.W.V.L. (2008). Detection of genetically modified organisms (GMOs) using molecular techniques in food and feed samples from Malaysia and Vietnam. International Food Research Journal 15(2): 155–166.
Yusuf, Z.K. (2010). Polymerase chain reaction (PCR). Saintek 5(6): 1–6.
Yuwono, T. (2010). Teori dan Aplikasi Polymerase Chain Reaction. Andi, Yogyakarta.
DOI: https://doi.org/10.22146/agritech.16656
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