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Journal : Makara Journal of Science

Optimization of pGEX System to Express and Isolate Mycobacterium tuberculosis Inclusion Body Protein in Combining with Modified Refolding Method Rukmana, Andriansjah; Burhanuddin, Burhanuddin; Yasmon, Andi
Makara Journal of Science Vol. 22, No. 4
Publisher : UI Scholars Hub

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Abstract

Antigen sub units for vaccine studies are typically isolated from recombinant proteins in an expression system. However, not all protein expression systems are used to express the specific protein. In this study,we optimized the pGEX system combined with the modified protein refolding to express and isolate M. tuberculosis proteins, especially proteins that are expressed as an inclusion body. Resuscitation promoting factor B (RpfB) protein is one of the Resuscitation promoting factor (Rpf) family of proteins that has been studied for its ability to induce cellular immunity in animal tests. Silico analyses demonstrate how RpfB is included in cell wall and cell processes. The Rpf family proteins are promising antigens that can be used as a TB vaccine candidate. The polymerase chain reaction was briefly performed using specific primers to amplify the full length of the rpfB. PCR amplification products were then purified, cut by restriction endonucleases, and cloned into pGEX 6-P1. Protein expression was done in the Escherichia coli BL21 strain, and expressed protein was isolated using themodified protein refolding and solubilization method. The complex protein expression that appeared as inclusion bodies were successfully isolated and can be detected as complex GST-RpfB through the western blotting process. Our study results indicate that this system and our modified method are suitable for M. tuberculosis inclusion body protein expression and isolation.
TROUBLESHOOTING IN EXPRESSION AND PURIFICATION OF RECOMBINANT SEVERE ACUTE RESPIRATORY SYNDROME-ASSOCIATED CORONAVIRUS NUCLEOCAPSID PROTEIN IN Escherichia coli BL21 Yasmon, Andi; Ibrahim, Fera; Bela, Budiman
Makara Journal of Science Vol. 14, No. 2
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Abstract

Co-Authors . Andriansjah Abror Irsan Ade P.R. Simaremare Agustini, Riani Anis Anis Anis Karuniawati Ardiana Kusumaningrum Ariyani Kiranasari ASRI SULFIANTI Beti Ernawati Dewi Budiman Bela Burhanuddin Burhanuddin Burhanuddin Burhanuddin Delly Chipta Lestari Delly Chipta Lestari Dewi Murniati Diah Dwi Utami Diana Natalia Diana Natalia Efendi, Ida Effendi, Ida Effiana Effiana Effiana, Effiana Elisabeth D. Harahap Elisna Syahruddin Erlina, Linda Fadilah Fadilah Fadilah FERA IBIRAHIM Fera Ibrahim Fithriyah Fithriyah Fithriyah Fithriyah Fithriyah Fithriyah Gultom, Desy A. Heri Wibowo Ibnu Agus Ariyanto Ika Ningsih Janah, Dariatul Junita Indarti Ketut Tuti Parwati Linosefa Linosefa Lisnawati Rachmadi Lola Febriana Dewi Louisa Ivana Utami Lucky H Moehario Mardhia, Mardhia Mardhia, Mardhia Mardiastuti H Wahid Maria L. Rosilawati Maria L. Rosilawati Maria Lina R. Maria Lina Rosilawati Maya Ulfah Moehario, Lucky Hartati Murti, Anissa Wari Ni Made Adi Tarini Ni Nengah Dwi Fatmawati Normasari Normasari Oktania Sandra Puspita Paramita, Rafika Indah Pramita G. Dwipoerwantoro Pratiwi Pujilestari Sudarmono Purba, Hastuti Handayani S Purnomosidi, Muhammad Abhi Rahayu, Ratih Ratnoglik, Suratno Lulut Rela Febriani Ria Kodariah Rifdah Hanifah Rosa, Yulia Rudyanto Sedono Safari, Dodi Salsabila, Korrie Samsuridjal Djauzi Sari Rahmayanti Simon Yosonegoro Liem SJATHA, FITHRIYAH Sudarmo, Fitrahwati Tafroji, Wisnu Teguh Sarry Hartono Thomas Robertus Tjampakasari, Conny Riana Utami, Diah Dwi VIVI SETIAWATY Winarti, Yayah Wresti Indriatmi B. Makes Wulandari, I Gusti Ayu Inten Yeva Rosana Yulia Rosa Saharman YULIANTY MUHAYAR Yusmaniar Yusmaniar Zaenab