Ekspresi gen hepatitis B surface antigen (sHBsAg) pada tanaman pisang transgenik untuk molecular farming vaksin

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Estiati, Amy, 2018, "Ekspresi gen hepatitis B surface antigen (sHBsAg) pada tanaman pisang transgenik untuk molecular farming vaksin", https://hdl.handle.net/20.500.12690/RIN/LPWOO7, RIN Dataverse, V1

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Description

Konsep baru yang berhubungan dengan vaksin adalah penggunaan tanaman sebagai pabrik untuk memproduksi vaksin. Permasalahan yang dihadapi adalah masih rendahnya tingkat ekspresi protein HBsAg dalam jaringan tanaman dibandingkan dengan sistem ekspresi HBsAg dari organisma inang lain. Penelitian yang telah dilakukan tim ini telah berhasil mengisolasi gen sHBsAg dari genom virus hepatitis B yang diperoleh dari serum pasien hepatitis B positif. Tujuan dari penelitian ini adalah mengembangkan vaksin edibel dengan melakukan transformasi gen Hepatitis B surface antigen (sHBsAg) kedalam tanaman pisang cv. Pisang Mas dengan bantuan A. tumefaciens, sehingga harapannya didapatkannya pisang transgenik mengandung gen sSHBsAg yang dapat diekspresikan. Prosedur dan metodologi penelitian terdiri atas transformasi gen sHBsAg, aklimitasi tanaman, analisis molekuler, isolasi DNA untuk PCR dan hibridisasi, analisis PCR, isolasi total RNA, reverse transcription PCR, PCR dan rapid kit test sHBsAg, uji hyromycin, SDS PAGE, ELISA, dan blotting. Hibridisasi Southern menggunakan probe berupa fragmen gen sHBsAg untuk mengetahui jumlah salinan gen dalam genom tanaman. Hanya tanaman dengan satu salinan gen saja yang akan dipilih untuk penelitian selanjutnya. Analisis ekspresi pada tingkat mRNA dan protein telah dilakukan untuk mengetahui keberadaan gen dalam tanaman. Hasil analisis PCR menunjukkan bahwa telah didapat 190 individu tanaman pisang terbukti mengandung gen sHBsAg dan gen hpt. Berdasarkan hasil RT-PCR diketahui 39 nomor tanaman mampu mengekspresikan gen sHBsAg dalam level mRNA dan dari hasil SDS- PAGE, ELISA dan Western blotting masing-masing diketahui 4 nomor, 1 nomor, dan 10 nomor berturut-turut mampu mengekspresikan gen sHBsAg dalam tingkat protein. Berdasarkan data analisis molekuler (analisis PCR, Rapid Kit Test HbsAg, SDS-PAGE, Western Blotting dan ELISA), disimpulkan bahwa nomor-nomor tanaman 52, A11, 341, 565, 358, 572, 371, 369, 341, 334, 337 adalah nomor-nomor potensial untuk dikonfirmasi menggunakan analisis molekuler kembali. Terdapat ekspresi HBsAg rekombinan pada level mRNA dan protein. Disarankan untuk melakukan hibridisasi Southern untuk menyeleksi tanaman dengan satu salinan gen sHBsAg. Perlu dilakukan analisis ekspresi gen sHBsAg dalam tanaman secara kuantitatif. Perlu dilakukan RT-PCR, Rapid Kit Test HbsAg, SDS-PAGE, Western Blotting dan ELISA untuk mendapatkan tanaman dengan ekspresi gen sHBsAg paling tinggi. (2011)

Subject

Agricultural Sciences


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Citation Metadata

Dataset Persistent ID	hdl:20.500.12690/RIN/LPWOO7
Publication Date	2018-02-16
Title	Ekspresi gen hepatitis B surface antigen (sHBsAg) pada tanaman pisang transgenik untuk molecular farming vaksin
Author	Estiati, Amy (Pusat Penelitian Bioteknologi LIPI)
Contact	Use email button above to contact. Estiati, Amy (Pusat Penelitian Bioteknologi LIPI)
Description	Konsep baru yang berhubungan dengan vaksin adalah penggunaan tanaman sebagai pabrik untuk memproduksi vaksin. Permasalahan yang dihadapi adalah masih rendahnya tingkat ekspresi protein HBsAg dalam jaringan tanaman dibandingkan dengan sistem ekspresi HBsAg dari organisma inang lain. Penelitian yang telah dilakukan tim ini telah berhasil mengisolasi gen sHBsAg dari genom virus hepatitis B yang diperoleh dari serum pasien hepatitis B positif. Tujuan dari penelitian ini adalah mengembangkan vaksin edibel dengan melakukan transformasi gen Hepatitis B surface antigen (sHBsAg) kedalam tanaman pisang cv. Pisang Mas dengan bantuan A. tumefaciens, sehingga harapannya didapatkannya pisang transgenik mengandung gen sSHBsAg yang dapat diekspresikan. Prosedur dan metodologi penelitian terdiri atas transformasi gen sHBsAg, aklimitasi tanaman, analisis molekuler, isolasi DNA untuk PCR dan hibridisasi, analisis PCR, isolasi total RNA, reverse transcription PCR, PCR dan rapid kit test sHBsAg, uji hyromycin, SDS PAGE, ELISA, dan blotting. Hibridisasi Southern menggunakan probe berupa fragmen gen sHBsAg untuk mengetahui jumlah salinan gen dalam genom tanaman. Hanya tanaman dengan satu salinan gen saja yang akan dipilih untuk penelitian selanjutnya. Analisis ekspresi pada tingkat mRNA dan protein telah dilakukan untuk mengetahui keberadaan gen dalam tanaman. Hasil analisis PCR menunjukkan bahwa telah didapat 190 individu tanaman pisang terbukti mengandung gen sHBsAg dan gen hpt. Berdasarkan hasil RT-PCR diketahui 39 nomor tanaman mampu mengekspresikan gen sHBsAg dalam level mRNA dan dari hasil SDS- PAGE, ELISA dan Western blotting masing-masing diketahui 4 nomor, 1 nomor, dan 10 nomor berturut-turut mampu mengekspresikan gen sHBsAg dalam tingkat protein. Berdasarkan data analisis molekuler (analisis PCR, Rapid Kit Test HbsAg, SDS-PAGE, Western Blotting dan ELISA), disimpulkan bahwa nomor-nomor tanaman 52, A11, 341, 565, 358, 572, 371, 369, 341, 334, 337 adalah nomor-nomor potensial untuk dikonfirmasi menggunakan analisis molekuler kembali. Terdapat ekspresi HBsAg rekombinan pada level mRNA dan protein. Disarankan untuk melakukan hibridisasi Southern untuk menyeleksi tanaman dengan satu salinan gen sHBsAg. Perlu dilakukan analisis ekspresi gen sHBsAg dalam tanaman secara kuantitatif. Perlu dilakukan RT-PCR, Rapid Kit Test HbsAg, SDS-PAGE, Western Blotting dan ELISA untuk mendapatkan tanaman dengan ekspresi gen sHBsAg paling tinggi. (2011)
Subject	Agricultural Sciences
Depositor	Puspita, Meilia
Deposit Date	2018-02-04
Related Datasets	Publikasi Gene transformation of hepatitis B surface antigen (HBsAg) on banana plants for molecuar farming vaccine. Estiati, Amy; Nurhasanah, Nena Ade; Witjaksono; Deswina, Puspita; Giri Rachman, Ernawati A.; Mulyana, Ima; Suhandono, Sony International Biotechnology Seminar, 2010 Introduction of hepatitis surface antigen (HbsAg) into banana cv. Mas Lampung for plant plant-based vaccine. Estiati, Amy; Nurhasanah, Nena Ade; Witjaksono; Deswina, Puspita; Giri Rachman, Ernawati A.; Mulyana, Ima; Suhandono, Sony Annales Bogorienses, 14(2) 2010: 21-25

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