Analisis Bioinformatika dan Ekspresi Protein Rekombinan Hemagglutinin Domain Globular dari Virus H5N1 Indonesia pada Eschericia coli BL21 (DE3) sebagai Komponen Vaksin Subunit Influenza


Muhammad Yusuf(1*), Rima Handiyani(2), Shinta Kusumawardani(3), Idar Idar(4), Umi Baroroh(5), Toto Subroto(6)

(1) Departemen Kimia, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Padjadjaran, Indonesia
(2) Departemen Kimia, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Padjadjaran, Indonesia
(3) Pusat Riset Bioteknologi Molekuler dan Bioinformatika, Universitas Padjadjaran, Indonesia
(4) Fakultas Farmasi, Universitas Bhakti Kencana, Indonesia
(5) Departemen Bioteknologi Farmasi, Sekolah Tinggi Farmasi Indonesia, Indonesia
(6) Departemen Kimia, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Padjadjaran, Indonesia
(*) Corresponding Author

Abstract


 

Flu burung merupakan salah satu penyakit zoonosis yang patut diwaspadai di Indonesia, khususnya galur High Pathogenic Avian Influenza (HPAI) karena dapat mematikan jika menular kepada manusia. Penggunaan vaksin influenza pada unggas, merupakan langkah preventif terhadap evolusi virus yang berbahaya dan juga penyebarannya. Selama ini, Indonesia masih menggunakan seed vaksin impor yang berasal dari luar Indonesia. Namun, karena Indonesia merupakan negara yang berada di garis khatulistiwa, karakteristik virus bisa berbeda dengan virus dari nothern-hemispere maupun southern-hemispere. Mengingat hal tersebut, Indonesia harus mengembangkan vaksin influenza menggunakan galur virus lokal. Berbeda dengan vaksin whole virus, vaksin rekombinan memiliki keunggulan dari sisi kemurnian, kecepatan produksi, dan kesesuaian galur terhadap virus yang beredar saat diperlukan. Penelitian ini bertujuan untuk menganalisis sekuen hemagglutinin (HA) Indonesia dengan strain lainnya serta mengekspresikkan protein HA1 rekombinan pada Escherichia coli  BL21 (DE3). Galur yang digunakan pada studi ini berasal dari virus H5N1 (A/Indonesia/05/05), khususnya bagian domain globular dari HA1. Sekuen HA1 bervariasi antara strain Indonesia dengan nothern-hemispere maupun southern-hemispere dan merupakan protein yang terpapar ke luar virus. Gen HA1 disisipkan pada vektor pET-28a, kemudian plasmid diisolasi menggunakan meoe manniatis, setelah itu diekspresikan dengan induksi 1 mM IPTG selama 4 jam. Protein HA1 telah berhasil diekspresikan secara intraseluler dan telah dikonfirmasi pada berat molekul 40 kDa menggunakan SDS-PAGE. Penelitian ini dapat digunakan untuk mengembangkan vaksin subunit yang lebih spesifik terhadap virus yang beredar di lapangan.


Keywords


Bioinformatika; influenza; hemagglutinin; protein rekombinan; Vaksin; Escherichia coli

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DOI: https://doi.org/10.15575/ak.v8i2.14138

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