IMUNOGEN UNTUK MENGHASILKAN ANTIBODI
SIMPULAN DAN SARAN
Simpulan
Berdasarkan hasil penelitian ini dapat disimpulkan bahwa gen SR100 berhasil diamplifikasi, kemudian diligasi dengan vektor pGEX-4T-2, dan ditransformasikan ke dalam bakteri E. coli DH5α. Hasil sekuensing menunjukkan
tidak terdapat mutasi pada gen hasil kloning. Plasmid pGEX-SR100 berhasil diekspresikan baik pada bakteri E. coli BL21 maupun E. coli DH5α. Ekspresi pada bakteri E. coli BL21 menghasilkan protein rekombinan lebih tinggi. Protein rekombinan HBsAg100-GST yang telah diekspresikan oleh E. coli tersebut berhasil dipurifikasi.
Saran
Penelitian dengan teknologi DNA rekombinan perlu digali dan dikembangkan terus dalam rangka pengembangan usaha industri peternakan khususnya di bidang penyediaan bahan vaksin. Pemanfaatan E. coli sebagai inang produksi protein rekombinan lebih tepat menggunakan E. coli BL21.
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73 Lampiran 1 Topologi dan peta fisik plasmid pGEM-T Easy
Sumber:
Diakses pada tanggal 13 Februari 2012 melalui:
http://www.promega.com/~/media/Files/Resources/Protocols/Technical%20Manu
75 Lampiran 2 (lanjutan)
77 Lampiran 2 (lanjutan)
Diterima melalui email dari Fak. Kedokteran UNRAM pada tanggal 2 April 2009
Lampiran 4 Topologi dan peta fisik plasmid pGEX-4T-2
79
81
Lampiran 5 (lanjutan)
Diakses pada tanggal 12 Juni 2011 melalui:
http://www.gelifesciences.com/aptrix/upp00919.nsf/Content/2CA907CE4753D32 BC1257628001D394F/$file/28918445AB.pdf
83 Lampiran 6 Hasil sekuensing gen SR100 dengan menggunakan primer pGEX-5’
Lampiran 6 (lanjutan)
85 Lampiran 7 Hasil sekuensing gen SR100 dengan menggunakan primer pGEX-3’
Lampiran 7 (lanjutan)
87 Lampiran 8 Mesin Thermal Cycler untuk mengamplifikasi segmen DNA
Lampiran 10 Alat elektroforesis (BIO-RAD) untuk memisahkan molekul protein berdasarkan berat molekulnya (tampak depan)
Lampiran 11 Alat elektroforesis (BIO-RAD) untuk memisahkan molekul protein berdasarkan berat molekulya (tampak atas)
89 Lampiran 12 Kelompok kandang mencit dalam penelitian
Lampiran 14 Proses mencampur HBsAg100-GST dengan Freund’s Adjuvant
sebagai bahan vaksin
91 Lampiran 16 Proses vaksinasi terhadap mencit sedang berlangsung
Lampiran 18 Hasil elisa dalam penentuan konsentrasi serum mencit untuk menguji antigenisitas protein rekombinan HBsAg100-GST
93 Lampiran 20 Printer yang terhubung dengan Mesin Elisa
Lampiran 21 Data hasil pembacaan optikal densiti (OD) terhadap serum mencit yang diperoleh dari darah mencit sebelum dan setelah dilakukan vaksinasi HBsAg100-GST (kelompok A) dan GST (kelompok B)