Assay of Bacillus subtilis ribonucleases in vitro.
Significant progress has been made recently regarding the identification of the ribonucleases involved in RNA maturation and degradation in Bacillus subtilis. More than half of these enzymes have no ortholog in Escherichia coli. To confirm that the in vivo effects of mutations in genes encoding RNases are direct, it is often necessary to purify the enzymes and assay their activity in vitro. Development of such assays is also necessary for detailed biochemical analysis of enzyme properties. In this chapter, we describe the purification and assay of 12 RNases of B. subtilis thought to be involved in stable RNA maturation or RNA degradation.
Condon
Ciarán
C
CNRS UPR 9073 (affiliated with Université de Paris 7; Denis Diderot), Institut de Biologie Physico-Chimique, Paris, France.
Pellegrini
Olivier
O
Mathy
Nathalie
N
Bénard
Lionel
L
Redko
Yulia
Y
Oussenko
Irina A
IA
Deikus
Gintaras
G
Bechhofer
David H
DH
D001412:Bacillus subtilis / Q000201:enzymology*; D001483:Base Sequence; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012341:RNA, Ribosomal, 5S / Q000737:chemistry; D012260:Ribonucleases / Q000302:isolation & purification / Q000378:metabolism*
D012341:RNA, Ribosomal, 5S; D012260:Ribonucleases