Isolation and cloning of the endoglucanase gene from Bacillus pumilus and its expression in Deinococcus radiodurans.
With the aim of engineering a strain of bacteria that could be used for bioremediation of cellulosic waste in radioactive environments, the gene for the secreted endoglucanase enzyme of Bacillus pumilis was decided to be cloned into the radiotolerant bacterium, Deinococcus radiodurans. The endoglucanase gene from B. pumilus was PCR amplified and cloned into Escherichiacoli DH5α using a pDrive vector. It was subsequently sub-cloned into E.coli-Deinococcus shuttle vector pRAD1 downstream of the Deinococcus heat-shock promoter, groESL, and the construct was inserted into D. radiodurans. Functional endoglucanase enzyme was expressed in both E.coli and D.radiodurans.
Department of Life Science and Biochemistry, Caius Research Laboratory, St. Xavier's College, Mumbai, 400001, India.|Caius Research Laboratory, St. Xavier's College, Mumbai, 400001, India.
Sachin
S
Telang
Caius Research Laboratory, St. Xavier's College, Mumbai, 400001, India.
Poonam
P
Patel
Caius Research Laboratory, St. Xavier's College, Mumbai, 400001, India.
Vishwas
V
Sarangdhar
Department of Life Science and Biochemistry, Caius Research Laboratory, St. Xavier's College, Mumbai, 400001, India. caiusreslab@gmail.com.|Caius Research Laboratory, St. Xavier's College, Mumbai, 400001, India. caiusreslab@gmail.com.|Indian Institute of Science Education and Research (IISER), Sai Trinity Building, Sutarwadi Road, Pashan, Pune, 411021, India. caiusreslab@gmail.com.
Sheela
S
Donde
Bacillus pumilus; Endoglucanase; groESL; pRAD1