Abstract
Publisher Summary 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 °.
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