Purification of ribonucleases Sa, Sa2, and Sa3 after expression in Escherichia coli.
The genes for three small ribonucleases from different strains of Streptomyces aureofaciens have been cloned and expressed in Escherichia coli. The purification of these ribonucleases from the periplasmic space is described. The yields range from 10 to 50 mg of protein per liter of culture medium. The molar absorption coefficients, isoelectric pH values, and pH of optimum activity are reported.
Hebert
E J
EJ
Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843, USA.
Grimsley
G R
GR
Hartley
R W
RW
Horn
G
G
Schell
D
D
Garcia
S
S
Both
V
V
Sevcik
J
J
Pace
C N
CN
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000302:isolation & purification; D012260:Ribonucleases / Q000235:genetics / Q000302:isolation & purification*; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology*
D007527:Isoenzymes; D011994:Recombinant Proteins; D012260:Ribonucleases; C077002:ribonuclease Sa3