Cloning and Expression of a Gene for Resistance to Butirosin both inEscherichia coliandBacillus subtilis

Abstract

We have cloned a butirosin resistance gene loc~ted on a 2.9 Md HindIII fragment of the chromosomal DNA of Bacillus vitellinus Z-1159, a butirosin producer, on a shuttle vector, pTA1302 (5.2Md, Apf, Cmf). The recombinant plasmid conferred resistances to butirosin, neomycin and kanamycin, on both Escherichia coli and Bacillus subtilis. Functional expression of aminoglycoside-3 '-phosphotransferase activity from the butirosin resistance gene was confirmed in the cell extracts of both E. coli and B. subtilis harboring the recombinant plasmid. A cleavage map of the recombinant plasmid was constructed.

Use of a Triple Protease-deficient Mutant ofBacillus subtilisas a Host for Secretion of aB. subtilisCellulase and TEMβ-Laetamase

“…plasmids, and culture conditions. B. subtilis RMI41leuBB arg15 recE4 hsrM hsmM 23 ) was used as a host for plasmid construction and B. subtilis 168 (a wild-type strain) and KN2 phe-Ilys-l trpC2 nprR2 nprEIB apr E!13 !1ispA 21) were used as hosts for secretion of BSC and BLA. Escherichia coli RRI supE44 hsdS20 (rB -mB -) ara-14 proA2 lacYI galK2 rpsL20 xyl-5 mtl-J24) and MV 1184 ara !1(lac-proAB) str A thi (4) Assay of cellulase activity.…”

Molecular Cloning of a Cellulase Gene fromBacillus subtilisand Its Expression inEscherichia coli

“…subtilis 1A274 npr, and B. subtilis RM141 leuB8, arg15, hsrM, hsmM, recE 4 , his 8 ) were used as hosts. B. subtilis IF03034 was the DNA donor for the cellulase gene.…”