Cloning and Expression of a Bacillus subtilis Endo-1,3-1,4- -D-Glucanase Gene in Escherichia coli K12
- 1 May 1984
- journal article
- research article
- Published by Microbiology Society in Microbiology
- Vol. 130 (5) , 1285-1291
- https://doi.org/10.1099/00221287-130-5-1285
Abstract
EcoRI fragments of DNA from B. subtilis NCIB 8565, a high producer of an endo-1,3-1,4-.beta.-D-glucanase, were shot-gun cloned in the plasmid vector pBR325. A 3.5 kb (kilobase) insert, carrying single restriction sites for AvaI, BglII, ClaI, PvuI and PvuII, was shown to direct the synthesis of .beta.-glucanase in E. coli K12. Enzyme activity was demonstrated in extracellular fractions of E. coli harboring the .beta.-glucanase gene. The largest proportion (> 50%) of total enzyme activity was periplasmic in location. .beta.-Glucanase activity and cellular location were independent of the orientation of the 3.5 kb fragment in pBR325.Keywords
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