Molecular cloning of an endoglucanase gene from an alkalophilic Bacillus sp. and its expression in Escherichia coli.
One of the cellulase genes from alkalophilic Bacillus sp. strain N-4 was cloned in pBR322. A recombinant plasmid, pYBC107, expressing carboxymethyl cellulase (CMCase) was isolated, and the size of the cloned HindIII fragment was found to be 5.5 kilobases. The restriction map of pYBC107 showed a different pattern from those of pNKI and pNKII (N. Sashihara, T. Kudo, and K. Horikoshi, J. Bacteriol. 158:503-506, 1984). When the HindIII fragment from pYBC107 was subcloned into pYEJ001, there was a 3.8-fold increase in CMCase activity over that observed with pYBC107. Plasmid pYBC108 constructed by treatment of pYBC107 with HindIII and EcoRI expressed the CMCase activity, although to a limited extent. To verify the originality of cloned pYBC107 from Bacillus sp., we analyzed the restriction digest by Southern blotting.
Kim
J M
JM
Department of Food Engineering, Yonsei University, Seoul, Korea.
Kong
I S
IS
Yu
J H
JH
D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D002480:Cellulase; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D015247:Deoxyribonuclease HindIII; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010957:Plasmids
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D015247:Deoxyribonuclease HindIII; D006026:Glycoside Hydrolases; D002480:Cellulase; C019826:carboxymethylcellulase