[Molecular cloning of cellulase gene from the bacillus]

The CMCase gene from the Bacillus sp. strain E2 was cloned into E. coli DH5 alpha F' using plasmid pBR322 as a vector. Recombinant plasmid pBG3 was isolated from transformants producing CMCase, and the cloned CMCase gene was found to be in a 4.0kb HindIII fragment. The CMCase gene was subcloned in a functional state on a 2.4kb DNA fragment. The optimum pH and temperature for activity of CMCase encoded by pBG3 were pH 6.5 and 55 degrees C respectively. The CMCase could not hydrolyze any natural cellulose examined. Eighty-three percent of the CMCase was detected in the inner cellular fraction, 9% and 8% were in the periplasmic space and medium respectively. Result of Southern hybridization indicates that the cloned DNA fragment was derived from Bacillus sp. strain E2 chromosomal DNA.