Exclusion chromatography with controlled-pore glass beads to isolate Chlorella chromatin and its applications.
A simple and rapid method was developed to isolate chromatin from the unicellular alga, Chlorella, by exclusion chromatography utilizing controlled-pore glass beads. This method takes advantage of the giant size of the chromatin supramolecules and does not require the preliminary isolation of cell nuclei. In order to raise the histone yield, commercially available materials were silanized with dimethyldichlorosilane. The isolated algal chromatin had properties similar to those of other organisms, and the histones contained all five components found in calf thymus. A hierarchy of the higher order structures was also observed in the algal chromatin. This method can be used for the study of chromatin in various cell types, especially in microbial cells, from the viewpoints of not only mere preparation but also cell dynamics and fractionation in relation to the specific components or activities. Some application examples are presented.
Fukumoto
T
T
Research Institute for Biochemical Regulation, Faculty of Agriculture, Nagoya University, Japan.
Nishimura
T
T
Nagasawa
T
T
Kitajima
K
K
Iwamura
T
T
D002453:Cell Cycle; D002708:Chlorella / Q000032:analysis* / Q000201:enzymology / Q000235:genetics; D002843:Chromatin / Q000096:biosynthesis / Q000302:isolation & purification*; D002850:Chromatography, Gel / Q000379:methods; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis; D004259:DNA-Directed DNA Polymerase / Q000032:analysis; D012321:DNA-Directed RNA Polymerases / Q000032:analysis; D005898:Glass; D006863:Hydrogen-Ion Concentration
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase