Isolation and characterization of the nuclease O gene (nucO) from Aspergillus oryzae.
Nuclease O in the mycelia of Aspergillus oryzae has been purified 55-fold by successive steps of chromatography from the filtrate of the autolyzate. The molecular mass of nuclease O was 32 kDa, as estimated by SDS polyacrylamide-gel electrophoresis. The nuclease O gene (nucO) encoding this enzyme was cloned and sequenced. The open reading frame is interrupted by four introns with conserved splice sites and contains 328 amino-acid residues of the mature enzyme. A. nidulans transformants obtained by introduction of the cloned nucO gene produced 2.5-times as much nuclease O as the wild-type strain, showing that the cloned DNA fragment encodes nuclease O.
Sano
M
M
Department of Biotechnology, Faculty of Agriculture, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan.
Nakamura
A
A
Masaki
H
H
Uozumi
T
T
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics; D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004706:Endodeoxyribonucleases / Q000235:genetics* / Q000302:isolation & purification*; D004722:Endoribonucleases / Q000235:genetics* / Q000302:isolation & purification*; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic
D004706:Endodeoxyribonucleases; D004722:Endoribonucleases; C021835:Serratia marcescens nuclease