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Journal : Makara Journal of Science

Cloning of a Gene Encoding Protease from Bacillus halodurans CM1 into Escherichia coli DH5α and Expression Analyses of the Gene Product Helianti, Is; Furgeva, Natasha; Mulyawati, Lina; Ferniah, Rejeki Siti; Kusumaningrum, Hermin Pancasakti
Makara Journal of Science Vol. 22, No. 3
Publisher : UI Scholars Hub

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Abstract

Bacillus halodurans strain CM1 is an Indonesia alkalothermophilic bacterium isolated from Cimanggu Hot Spring, Bandung, West Java. This bacterial strain produces high levels of thermoalkalophilic xylanase. It has also been predicted to produce other potential industrial enzymes, including protease. For production and application of protease in the future, the protease gene from B. halodurans CM1 was cloned into Escherichia coli. The protease gene was isolated from B. halodurans CM1 by the PCR approach using primers designed based on the GenBank. The PCR product was then ligated into pGEM-T Easy vector, transformed into E. coli DH5α, verified, and analyzed based on DNA sequencing data using the BLAST search tool. A 1086-bp protease gene was obtained that exhibited a very high sequence similarity (99%) with that of alkaline protease gene from B. halodurans C-125. When the culture of this positive recombinant E. coli DH5α containing the protease gene was spotted onto calcium caseinate agar, a clear zone appeared after incubation at 50 °C. This result demonstrated that the protease gene was expressed in this recombinant E. coli DH5α.
Co-Authors Agung Suprihadi Agung Suprihadi Agus Sabdono Agus Setiadi Agus Subagio Anggoro, Naufal Sebastian Annisa Fadillah Annisa Nur Ayuningtyas Anto Budiharjo Arina Tri Lunggani Aris Ismanto Asih Rismiarti, Asih Athried Elsima Azis Rifai Baskoro Rochaddi Bintoro Rudi Saputro, Bintoro Rudi Budi Raharjo Chatarina Umbul Wahyuni Choirunnisaa, Nadia Maharani Jasmine Christina Ratna Handayani, Christina Ratna Christina Retna Handayani, Christina Retna Clara Yully Diana Ekaristi Denis Denny Nugroho Sugianto Denny Nurkertamanda Dinda Khairunnisa, Dinda Doktorasaintifika, Heradita Kaniaazzahra Dudi Hardianto, Dudi Eiffeliena Nur'aini Fisikaningputri Purwienanti Eko Purnomo Elke Gildantia Elsima, Athried Endang Dwi Purbajanti Endang Kusdiyantini Erfianti, Tia Fiona Aqhila Dewi Fitri Arum Sasi, Fitri Arum Furgeva, Natasha Gentur Handoyo Hadi Endrawati Hapsari, Hererapratita Aysha Heksa Raihan Hererapratita Aysha Hapsari Herida, Azalia Puspa Hersugondo Hersugondo Hersugondo Hersugondo Herusugondo Herusugondo Imam Misbach Imam Mishbach Indah Sulistyarini Indras Marhaendrajaya Is Helianti IS HELIANTI Istiana, Rohma Ita Widowati Izzudin, Maulana Zidan Jihadi, Muhammad Shulhan Joedono Soedarsono Joedoro Sudarsono Karima, Radhita Kurniawati, Mufida Budi Lilis Sugiarti Lilis Sugiarti Lina Mulyawati Maria, Atina Mawarni, Shelina Nurunnisa Maya Aresteria MG Isworo Rukmi Monalita, Ramadhebi Muhammad Amal Nurhakim, Muhammad Amal Muhammad Anwar Djaelani Muhammad Iskandar Zulkarnain Muhammad Iskandar Zulkarnain Muhammad Mulyadi, Muhammad Muhammad Zainuri Mulyawati, Lina Natasha Furgeva Nurhayati Nurhayati Nurhayati Nurhayati Okti Hajeng Kristiadi Rahayu, Annisa Rizky Rejeki Siti Ferniah Retno Hartati Risma Wiharyanti Rizko, Nurmalisa Robertus Triaji Mahendrajaya Roslenawati Roslenawati Rudhi Pribadi Safirah, Dearesty Sarwo Edi Wibowo, Sarwo Edi Shasabila, Analies Devi Siti Nur Jannah Siti Nur Jannah Solly Aryza Sonny Abdi Setiyawan, Sonny Abdi Sri Pujiyanto Sri Pujiyanto Sri Pujiyanto Sri Pujiyanto Sri Rustini Subagiyo Subagiyo Sulis Setyowati, Sulis Suryani, Oda Gracia Ariela Sutrisno Anggoro Tony Hadibarata, Tony Tri Rahayu, Hesti Triwibowo Yuwono Wahyu Dewi Utari Haryanti Wahyuningsih, Candra Widianingsih Widianingsih Wijanarka Wijanarka Yann Hardivillier Yason Lukman Sudjito, Yason Lukman Yosef Purwoko Yoshua Mario Sumbodo Yudi Yunanto Yumna Rahmadias Hanifa Yunior William Susanto Yuvita Muliastuti