Rekayasa Bakteri untuk Ternak dan Manusia: Pembuatan Mutan Escherichia coli Penghasil Protein Rekombinan

Muhamad Ali, Takako FUKUBA, Hideo NAKANO

Abstract


Protein rekombinan seperti vaksin, antibodi, hormon, dan obat-obatan, semakin dibutuhkan oleh ternak dan manusia. Hambatan utama untuk menghasilkan protein rekombinan pada Escherichia coli sebagai inang yang digunakan paling luas adalah degradasi oleh enzim proteolitik. Hal ini disebabkan karena E. coli memiliki sejumlah enzim proteolitik yang tersebar di dalam sitoplasmanya. Untuk itu, lebih dari 90% degradasi protein terjadi di dalam sitoplasmanya. Pada penelitian ini, peneliti telah menghasilkan mutant E. coli BW25113 yang tidak memiliki gen penyandi enzim protease dengan menggunakan kombinasi metode pengerusakan kromosom dan metode transduksi phage P1. Pembuatan mutan tersebut dimulai dengan pengerusakan gen penyandi enzim protease pada kromosom bakteri dengan produk PCR yang memiliki bagian yang homolog dengan gen target. Mutan-mutan yang dihasilkan kemudian digunakan untuk menghasilkan mutan ganda dengan metode Transduksi phage P1. Analisis fenotif dan genotif menunjukkan bahwa kombinasi kedua metode tersebut sangat efektif untuk membuat lebih dari satu mutasi pada E. coli. Untuk itu, mutan E. coli yang telah diperoleh akan sangat bermanfaat untuk menghasilkan aneka protein rekombinan untuk ternak dan manusia.

Keywords


mutan defisiensi-protease; transduksi phage P1; disrupsi kromosom

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DOI: https://doi.org/10.33476/jky.v18i2.184

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