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All Journal HAYATI Journal of Biosciences VALENSI Bionatura Microbiology Indonesia Majalah Kedokteran Bandung Jurnal Riset Kimia Indonesian Journal of Pharmaceutical Science and Technology JC-T (Journal Cis-Trans): Jurnal Kimia dan Terapannya Health Notions Chimica et Natura Acta ALCHEMY Jurnal Penelitian Kimia Jurnal Kimia Terapan Indonesia Jurnal Pendidikan Kimia (JPKim) Indonesian Journal of Chemistry PENDIPA Journal of Science Education Molekul: Jurnal Ilmiah Kimia Jurnal Ilmiah Farmako Bahari
Shabarni Gaffar
Departemen Kimia, Fakultas Matematika Dan Ilmu Pengetahuan Alam, Universitas Padjadjaran
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Dentistry Earth & Planetary Sciences Education Energy Environmental Science Health Professions Immunology & microbiology Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Nursing Public Health

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Journal : microbiology indonesia

 1 
Codon Optimization and Chaperone Assisted Solubilization of Recombinant Human Prethrombin-2 Expressed in Escherichia coli SARONOM SILABAN; IMAN PERMANA MAKSUM; SHABARNI GHAFFAR; KHOMAINI HASAN; SUTARYA ENUS; TOTO SUBROTO; SOETIJOSO SOEMITRO
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 (692.802 KB) | DOI: 10.5454/mi.8.4.5

Abstract

Prethrombin-2 (PT2) is a thrombin precursor, which plays a role in the conversion of fibrinogen into fibrin during blood clotting process. Previous study reported that the expression of human prothrombin-2 (rhPT2) in Escherichia coli formed inclusion bodies. The aim of this study was to establish a strategy to express a soluble rhPT2 in E. coli. This study was animed to design and codon optimize human prethrombin-2 gene as well as to optimize the expression condition using four strains of E. coli. The codon adaptation index (CAI) of the unoptimized hpt2 gene was 0.336, with 56.8% GC content. After optimization, the CAI of optimized hpt2 became 1.000 with 53.1% GC content. The optimized gene was successfully cloned into pTWIN1 expression vector. Expression analysis indicated that only E. coli ArcticExpress strain could successfully express a soluble recombinant rhPT2 protein, with only part of rhPT2 being expressed in insoluble form. However, the rest of the E. coli strains used in the experiments failed to express the rhPT2 in soluble form. We are deducing that the success in achieving soluble expression was not only due to the availability of chaperonins Cpn60/Cpn10, which played a crucial role in the protein folding in E. coli ArcticExpress strain, but also due to the codon optimization of hpt2 gene.
Co-Authors Aathirah, A Sayyidatina Abu Bakar M.I. Syihab Ade Rizki Ridwan Firdaus Aga Adi Masyhuri Agus Safari Al Arofatus Naini Albayyinah, Dyandra Hera Amalia, Riezki Anni Anggraeni, Anni Annisa Ilma Naviardianti Annisa Ilma Naviardianti Ari Hardianto Arum Kurnia Sari Bacthi Alisjahbana Bayu Shiddiq Widhi Pratama Dani Permana Darwati Darwati Desi Harneti Putri Huspa DESSY NATALIA Diana P Rahmawati Egista Istioka Fazrin Ekawardani, Savira Endang Srieatimah - Erina Hilmayanti Ersanda Hafiz Euis Julaeha Euis Julaeha Fajriyah, Maulida Ghina Nur Fadhilah Ghina Uli Felicia Tambunan Hafiz, Ersanda Hesti Lina Wiraswati Iman Permana Maksum Iqbal Wahyu Mustaqim Irkham Irkham KHOMAINI HASAN Kindi Farabi Korry Novitriani, Korry Lia Faridah LINAWATI HARDJITO Lubis, Rubianto A. M. Lutfi Firdaus Ma'ruf, Ilma Fauziah Masyhuri, Aga Adi Mohamad Nurul Azmi Muhammad Hilman Daniswara Muhammad Yudha Nugraha Muhammad Yusuf Murniaty Simorangkir Nayla Haraswati Nurlelasari Nurlelasari Nurmalasari, Ratna Nurul Auliany O Suprijana Oo Suprijana Permadi, Nandang Pertiwi, Wulan Purba Upay Purnama Purnama PUTRI, RAFIKA Putri, Rafika Nanda R. Ukun M.S. Soedjanaatmadja Rani Maharani Rima Melati Rini Surbakti Riza Apriani Riza Apriani Riza Apriani Rubianto A. Lubis Rudi Hartono Rustaman Rustaman Saadah D. Rachman Safri Ishmayana Safrl Ismayana Santhy Wyantuti Sari Syahruni SARONOM SILABAN Saronom Silaban Savira Ekawardhani Shabrinna, Hanif SOETIJOSO SOEMITRO Soetijoso Soemitro Soetijoso Soemitro Sofa Fajriah Sofyan Multazam N Aji Supriyadi, Isma Yustifania SUTARYA ENUS Sutarya Enus Syifa Al Fauziah Rahmani Tambunan, Ghina Uli Felicia Tati Herlina Tati Herlina Toto Subroto Tri Mayanti Triana N Meirina Umi Baroroh Unang Supratman Upay, Purba Vicki Nishinarizki Wulan Pertiwi Yeni W Hartati Yeni Wahyuni Hartati Yenni Wahyuni Hartati, Yenni Wahyuni Yohan Yohan Yohan Yohan Yoshihito Shiono