TY - JOUR
T1 - [13] Purification and Properties of HindII and HindIII Endonucleases from Haemophilus Influenzae Rd
AU - Smith, Hamilton O.
AU - Marley, Garry M.
N1 - Funding Information:
Work performed in the authors' laboratory was supported by grants from the National Institutes of Health and the National Science Foundation.
PY - 1980/1/1
Y1 - 1980/1/1
N2 - This chapter discusses the purification and properties of HindII and HindIII endonucleases from Haemophilus influenzae Rd. Haemophilus influenzae Rd is the source for two restriction endonucleases,HindII and HindlII, that may be separated and purified using conventional ion exchange chromatography media. The basic steps in the procedure are removal of nucleic acids from the cell extract, separation of the two activities by DEAE-cellulose chromatography, and chromatography of each activity on phosphocellulose as a combined purification and concentration step. The chapter also presents separation of HindlI and HindllI activities by DEAE-cellulose chromatography. In addition, properties are also overviewed, such as enzyme stability. HindlII is stable for at least 2 years at -20 °. Under reaction conditions it retains activity for hours at 37 ° and for several minutes at 65 °.
AB - This chapter discusses the purification and properties of HindII and HindIII endonucleases from Haemophilus influenzae Rd. Haemophilus influenzae Rd is the source for two restriction endonucleases,HindII and HindlII, that may be separated and purified using conventional ion exchange chromatography media. The basic steps in the procedure are removal of nucleic acids from the cell extract, separation of the two activities by DEAE-cellulose chromatography, and chromatography of each activity on phosphocellulose as a combined purification and concentration step. The chapter also presents separation of HindlI and HindllI activities by DEAE-cellulose chromatography. In addition, properties are also overviewed, such as enzyme stability. HindlII is stable for at least 2 years at -20 °. Under reaction conditions it retains activity for hours at 37 ° and for several minutes at 65 °.
UR - http://www.scopus.com/inward/record.url?scp=0018818001&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0018818001&partnerID=8YFLogxK
U2 - 10.1016/S0076-6879(80)65015-0
DO - 10.1016/S0076-6879(80)65015-0
M3 - Article
C2 - 6246330
AN - SCOPUS:0018818001
SN - 0076-6879
VL - 65
SP - 104
EP - 108
JO - Methods in enzymology
JF - Methods in enzymology
IS - C
ER -