MOLECULAR EXPRESSION OF WINGLESS-TYPE MMTV INTEGRATION SITE FAMILY MEMBER 4 GENE USING Escherichia coli BL21

Jurnal Kedokteran Hewan, Agung Sitasiwi, Wayan Tunas Artama, Agung Budiyanto, Edy Dharmana
2017 unpublished
This research was conducted to find out the Wnt4 recombinant proteins which expressed by Escherichia coli (E. coli) BL21 carrying the recombinant DNA wnt4 (E. coli transformation). Research materials were E. coli BL21 transformation and E. coli BL21 non-transformation (negative control). The expression of recombinant protein was conducted by culturing E. coli for 24 hours in Luria-Bertani (LB) media with isopropyl β-D-1-thiogalactopyranoside (IPTG) induction. Recombinant protein was isolated by
more » ... ein was isolated by sonication of pellet bacteria. Protein analysis performed by 15% sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that recombinant protein with a molecular weight of 33 kDa has been expressed by E. coli BL21 transformation successfully. ____________________________________________________________________________________________________________________ ABSTRAK Penelitian ini dilakukan dengan tujuan mendapatkan protein rekombinan Wnt4 yang diekspresikan oleh Escherichia coli (E. coli) BL21 pembawa deoxyribonucleic acid (DNA) rekombinan wnt4 (E. coli transforman). Materi uji ekspresi protein adalah E. coli BL21 transforman dan E. coli BL21 nontransforman (sebagai kontrol negatif). Ekspresi protein rekombinan dilakukan dengan cara menumbuhkan bakteri selama 24 jam pada media Luria-Bertani (LB) cair dengan induksi isopropyl β-D-1-thiogalactopyranoside (IPTG). Protein rekombinan diisolasi dengan cara sonikasi pelet bakteri. Analisis protein dilakukan dengan sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) dengan konsentrasi 15%. Hasil penelitian menunjukkan bahwa protein rekombinan dengan berat molekul 33 kDa berhasil dieskpresikan oleh E. coli BL21 transforman. ____________________________________________________________________________________________________________________ Kata kunci: E. coli BL21, imunokontrasepsi, wnt4
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