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- Title
Cloning of DNA Polymerase I Geobacillus thermoleovorans SGAir0734 from a Batu Kuwung Hot Spring in Escherichia coli.
- Authors
Lischer, Kenny; Tansil, Kevin Priyono; Ginting, Mikael Januardi; Sahlan, Muhamad; Wijanarko, Anondho; Masafumi Yohda
- Abstract
Access to biological engineering has become a critical point of modern science development through polymerase chain reaction (PCR). One of the main components in this process is DNA polymerase, which copies the main template DNA. However, there is a lack of studies on the production of DNA polymerase from indigenous thermophilic bacteria in Indonesia. To examine this process, DNA polymerase I gene (DNA pol I) from Geobacillus thermoleovorans (isolated from Batu Kuwung, Banten, Indonesia) was transformed into Escherichia coli. The gene was cloned by the cut and ligation method using NcoI and BamHI restriction enzymes, which were ligated with a pET23d vector. The recombinant gene was overexpressed in E. coli and identified by using SDS-PAGE of 10% acrylamide gel, showing that the protein molecular weight was approximately. This study successfully amplified the gene of interest, indicated by a high local similarity between the sequencing results and theoretical gene and positive intercellular protein expression. The results confirm that the study successfully cloned and synthesized recombinant DNA pol I of Geobacillus thermoleovorans from Batu Kuwung, Serang, Banten.
- Subjects
INDONESIA; MOLECULAR cloning; DNA polymerases; HOT springs; RECOMBINANT DNA; ESCHERICHIA coli; BIOENGINEERING
- Publication
International Journal of Technology, 2020, Vol 11, Issue 5, p921
- ISSN
2086-9614
- Publication type
Article
- DOI
10.14716/ijtech.v11i5.4311