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All Journal MANAJEMEN HUTAN TROPIKA Journal of Tropical Forest Management METAMORFOSA Journal of Biological Sciences Biotika: Jurnal Ilmiah Biologi Bioscience Microbiology Indonesia Jurnal Pendidikan Sains AL KAUNIYAH Jurnal Sains dan Teknologi Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Biosfer: Jurnal Tadris Biologi Biogenesis: Jurnal Ilmiah Biologi Quagga Gontor AGROTECH Science Journal Biology, Medicine, & Natural Product Chemistry Journal of Natural Sciences and Mathematics Research Jurnal Biodjati Jurnal Pengabdian Pada Masyarakat JRST (Jurnal Riset Sains dan Teknologi) Biosel: Biologi Science and Education Jurnal Biolokus: Jurnal Penelitian Pendidikan Biologi dan Biologi KACANEGARA Jurnal Pengabdian pada Masyarakat AL-HAYAT: Journal of Biology and Applied Biology Logista: Jurnal Ilmiah Pengabdian Kepada Masyarakat Jurnal Pemberdayaan: Publikasi Hasil Pengabdian Kepada Masyarakat Kontribusia : Research Dissemination for Community Development International Journal of Engineering, Technology and Natural Sciences (IJETS) Jurnal Berdaya Mandiri Jurnal Sains dan Edukasi Sains Indonesian Journal of Applied Research (IJAR) Indobiosains Bioscientist : Jurnal Ilmiah Biologi Jurnal Pengabdian Masyarakat Indonesia International Journal of Research in Community Services Aptekmas : Jurnal Pengabdian Kepada Masyarakat International Journal on Education Insight Journal of Biotechnology and Natural Science Filogeni: Jurnal Mahasiswa Biologi Jurnal Pengabdian Masyarakat Bangsa DIMASLIA Jurnal Abdimas
Oktira Roka Aji
Program Studi Biologi Fakultas Sains Dan Teknologi Terapan Universitas Ahmad Dahlan
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Journal : Microbiology Indonesia

 1 
Cloning, Overexpression, and Purification of PhoR CytoplasmicDomain Protein from Mycobacterium tuberculosis strain H37Rv OKTIRA ROKA AJI; DYSHELLY NURKARTIKA PASCAPURNAMA; FENRYCO PRATAMA; IHSANAWATI IHSANAWATI; MAELITA RAMDHANI MOEIS; ERNAWATI ARIFIN GIRI-RACHMAN
Microbiology Indonesia Vol. 8 No. 4 (2014): December 2014
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (758.166 KB) | DOI: 10.5454/mi.8.4.1

Abstract

Tuberculosis still becomesa major health problem in the world. This infectious disease is caused by Mycobacterium tuberculosis (Mtb). Novel anti-tubercular drug is urgently needed to counter multidrug resistant cases and Mtb's spread. The cytoplasmic domain of PhoR histidine kinase, a part of the two-component system PhoR-PhoP in Mtb, is one of the potential candidates for anti-tubercular drug target.Three dimensional protein of drug target is needed to screen potential drug candidate using rational drug design approaches. Previous studies have successfully characterized and isolated putative cytoplasmic domain of PhoR (CytoPhoR) from Mtb strain H37Rv. This study aimed to clone, overexpress and purify of CytoPhoR protein. CytoPhoR was fused with thioredoxin protein in expression vector pET32b and overexpressed in Escherichia coli (E.coli)BL21 (DE3) as soluble fraction by induction  1 mM IPTG. Purification of his-tagged CytoPhoR was carried out using IMAC Ni-NTA Agarose his-tag affinity column. SDS-PAGE analysis showed that another protein was co-purified (~35 kDa) along with the CytoPhoR protein. Subsequent protein purification using DEAE-ion exchange column generate a strong single band of 37 kDa on SDS–PAGE which is indicated as CytoPhoR protein. The purified CytoPhoR protein was successfully obtained and can be used for further analysis on determining three dimensional structure of CytoPhoR protein.
Expression and Purification of PhoR Sensor-Domain Histidine Kinase of Mycobacterium tuberculosis in Escherichia coli ERNAWATI ARIFIN GIRI-RACHMAN; FENRYCO PRATAMA; OKTIRA ROKA AJI; ARUM PATRIATI; IHSANAWATI IHSANAWATI; MAELITA RAMDANI MOEIS; EDY GIRI-RACHMAN PUTRA
Microbiology Indonesia Vol. 9 No. 2 (2015): June 2015
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1382.183 KB) | DOI: 10.5454/mi.9.2.1

Abstract

Globally, tuberculosis (TB) remains a leading cause of death. The emergence of multidrug-resistant strains (MDR-TB) and extensively drug-resistant strains (XDR-TB) has fuelled the discovery for novel drugs and drug targets for its successful and better treatment. One of the potential candidates for drug target is PhoR sensory protein histidine kinase, a part of the Two Component System (TCS) PhoP/PhoR in Mycobacterium tuberculosis (Mtb). This protein system was known for its role on regulating hundred of Mtb virulence factors, from genes for cell wall and lypid synthesis to genes for adaptation in human leukocyte and hypoxia response. Previous studies have successfully characterized, isolated, and cloned the putative sensory domain of PhoR protein gene into pRSET vector expression system. In this study, Escherichia coli was transformed with pRSET-SensPhoR and cultivated at 37oC under IPTG induction to express PhoR sensor-domain protein. Most of the proteins were overexpressed in the form of inclusion bodies.  Subsequent protein purification in Ni-NTA system under refolding condition on urea gradient was performed to isolate PhoR sensor-domain protein in soluble form. Arginine was supplemented in purified protein solution to prevent aggregation during long term storage.  While highly purified protein was acquired, small angle X-ray scattering (SAXS) analysis was conducted to obtain 3-dimensional (3D) protein structures in solution.    doi:10.5454/mi.9.2.1 
Co-Authors Abdul Aziz Adz-Dzikir, Fatihatuljannah Aila Gema Safitri Aisyah, Iis Nisya Nur Al Farisi, Irsyad Andika Bayu Chandra Aninda Kumala Sari Anisa Nurul Huda Ardi, Subhan Zul Arifa Nadya Nur Afifah ARUM PATRIATI Arumbina, Vibula Rosa Astuti, Sutan Nur Chamida Tri Azizah, Mar'atul Azizah, Rara Asti Cahya Ningsih, Ayu Cucu Cahyanti Diah Asta Putri Diah Asta Putri Diah Asta Putri Diah Asta Putri DYSHELLY NURKARTIKA PASCAPURNAMA EDY GIRI-RACHMAN PUTRA Efrida Siti Alifia Ega Asnatasia Maharani ERNAWATI ARIFIN GIRI-RACHMAN Falih, Afifah Nurul FENRYCO PRATAMA Fitria, Karomah Indah Handayani, Erisa Budi Hartanto, Dody Haryadi, Bagus Hasna Chaerani Zakkiyah Hidayah, Hidayah Hudaya, Riris Asyhar IHSANAWATI IHSANAWATI Ihsani, Nisa Ika Maryani Ilma, Nur Malika Imelda Pangaribuan Inggita Utami Irsalinda, Nursyiva Irvanita, Deva Anggun Jannati Aulah Jinan, Rifa Fatma Kamilah, Umu Kurnia, Dhiffa Trisna Laila, Alfiatul Larasati Haliimah Roosyidah Lestari, Iva Dita Lestari, Kinasti Dwi Linda Pertiwi MAELITA RAMDANI MOEIS MAELITA RAMDHANI MOEIS Maliza, Rita Mardiah, Irma Merlia Rahma Tari Muhammad Farid Maksum Nisa Ihsani Nofa Nur Fiani Nora Bastiani Nugraha, Aisyah Aulia Nugraha, Farhan Satya Nur Azizah Nur Azizah Nuraisyah Binte Zainal Nurul Suwartiningsih Pinasti, Vanya Aurel Pratiwi, Ambar Pratiwi, Ambar Puguh Wahyu Prasetyo, Puguh Purwanti Pratiwi Purbosari Puspitarini, Sofia Putri , Diah Asta Putri, Anas Tasya Putri, Aulia Ramanda Putri, Diah Asta Putri, Nida’a Fauziyyah Rinaldi Rizal Putra Riris Asyhar Hudaya Rohmantin, Dita Rossa Puteri Baharie, Sri Safitri, Salsabila Salimah, Azizah Zahratus Santrianda, Aldhiani Setiawan, Haris Silfi Pratiwi Sri Wahyuni Sri Wahyuni Sumbudi, Milliniawati Suwartiningsih, Nurul Suwartiningsih, Nurul Tiwi Sartika Tri Puspita Sari Tufahati, Nadhira Umi Salamah Vianingtyas, Naresh Wari Mega Wachid Eko Purwanto Wicaksono, Adelyanty Pangestuty Wulandari, Endah Retno Yudi Ari Adi, Yudi Ari Yuliani, Ririn Kholifatu Zahra, Safa Auli Zakkiyah, Hasna Chaerani Zul Ardi, Subhan