Konstruksi Plasmid pET-15b dengan Gen tat Virus Penyakit Jembrana

Asmarani Kusumawati; Lalu Unsunnidhal; Agung Budiyanto; Erif Maha Nugraha Setyawan; Sri Gustari

doi:10.22146/jsv.79217

Konstruksi Plasmid pET-15b dengan Gen tat Virus Penyakit Jembrana

https://doi.org/10.22146/jsv.79217

Asmarani Kusumawati^(1*), Lalu Unsunnidhal⁽²⁾, Agung Budiyanto⁽³⁾, Erif Maha Nugraha Setyawan⁽⁴⁾, Sri Gustari⁽⁵⁾

(1) Departemen Reproduksi dan Kebidanan Fakultas Kedokteran Hewan Universitas Gadjah Mada
(2) Program Studi Ilmu dan Teknologi Pangan Fakultas Teknologi Pangan dan Agroindustri Universitas Mataram
(3) Departemen Reproduksi dan Kebidanan Fakultas Kedokteran Hewan Universitas Gadjah Mada
(4) Departemen Reproduksi dan Kebidanan Fakultas Kedokteran Hewan Universitas Gadjah Mada
(5) Departemen Reproduksi dan Kebidanan Fakultas Kedokteran Hewan Universitas Gadjah Mada
(*) Corresponding Author

Abstract

Virus Penyakit Jembrana adalah virus yang menyebabkan jembrana pada sapi bali. Vaksin protein rekombinan dibuat dengan memasukkan DNA atau gen yang mengkode protein imunogenik ke dalam vektor ekspresi prokariotik sehingga dapat mengekspresikan protein rekombinan yang akan dijadikan vaksin. Salah satu gen struktural virus penyakit jembrana adalah gen tat yang mengekspresikan protein tat yang digunakan sebagai vaksin protein rekombinan. Tujuan dari penelitian ini adalah untuk mengkloning pET15b dengan gen tat. Konfirmasi gen pET-15b pada bakteri transforman dilakukan dengan koloni PCR, kemudian dilakukan isolasi plasmid dan dilaksanakan konfirmasi menggunakan metode PCR dengan primer untuk deteksi gen tat dan primer sekuensing untuk vektor pET15b. Hasilnya menunjukkan bahwa bakteri transforman sudah berisikan DNA rekombinan pET15b-tat yang terkonfirmasi melalui koloni PCR dengan band sebesar 179 bp, kemudian dikonfirmasi lanjut melalui PCR pada hasil isolasi plasmid dari bakteri transforman yang didapatkan band sebesar 179 bp untuk primer deteksi gen tat dan 514 bp untuk primer sekuensing pada vektor pET15b, 179 bp dan 514 bp sesuai dengan target amplikasi untuk primer deteksi dan primer sekuensing yang digunakan.

Keywords

Virus Penyakit Jembrana; Vaksin Protein Rekombinan; Protein tat; pET15b; PCR

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References

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DOI: https://doi.org/10.22146/jsv.79217