Dye-ligand affinity chromatography of RNA polymerase II.
The binding of wheat germ RNA polymerase II to five different dye-ligand chromatography gels (Matrex gels, Amicon Corp.) was tested. A quantitative binding of the enzyme to four of the gels, namely Dyematrex gels Blue A, Blue B, Red A and Green A was observed. Only the Orange A gel column failed to bind the enzyme strongly. Nearly 100% of the activity could be recovered from the Green A column by elution with high salt concentration and high pH. Under these conditions only a part of the activity was eluted from the other three columns since the enzyme bound tightly. Enzyme activity could be removed from the columns by elution with nucleotide substrates, but the yield from the Blue A, Blue B and Red A columns was still low (7 to 42%). The Green A Matrex gel appeared to be useful for the purification and analysis of RNA polymerase.
Skripal, IG; Weeks, JR; Greenleaf, AL
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