Cloning the hbs gene from Bacillus subtilis and expression of the HBsu protein in Escherichia coli
Ghodsi، S نويسنده Biology Department, Faculty of Sciences, Alzahra University, Vanak, Tehran, IR Iran. Ghodsi, S , Gharavi، S نويسنده Biology Department, Faculty of Sciences, Alzahra University, Vanak, Tehran, IR Iran. Gharavi, S , Ghadam، P نويسنده Biology Department, Faculty of Sciences, Alzahra University, Vanak, Tehran, IR Iran. Ghadam, P
Background and Objectives: Bacillus subtilis HBsu is a 10 kD heat-stable protein shown to be involved in binding to DNA and is encoded by the hbs gene. Large-scale production for biochemical analysis is achieved through cloning and expression of the recombinant protein.
Materials and Methods: This gene was amplified from B. subtilis ATCC 6633 using PCR and cloned into pET28a (+) expression vector. The construct was used to transform Escherichia coli BL21 (DE3). The expression of the protein was induced by the addition of 1mM IPTG. To confirm the expression of the cloned gene, SDS-PAGE was carried out and production of an approximately 11 KD recombinant tagged protein was confirmed for the cloned hbs gene.
Results and Conclusion: The identity of the recombinant HBsu was verified and characterized by SDS-PAGE which can then be utilized for further applications.