PURIFICATION AND CHARACTERIZATION OF THE M.RSRI DNA METHYLTRANSFERASE FROM ESCHERICHIA-COLI

被引：22

作者：

KASZUBSKA, W ^{[1
]}

WEBB, HK ^{[1
]}

GUMPORT, RI ^{[1
]}

机构：

[1] UNIV ILLINOIS,COLL MED,DEPT BIOCHEM,415 RAL,1209 W CALIF ST,URBANA,IL 61801

来源：

GENE | 1992年 / 118卷 / 01期

关键词：

DNA METHYLATION; FLUORESCENCE SPECTROSCOPY; VELOCITY SEDIMENTATION; SINEFUNGIN;

D O I：

10.1016/0378-1119(92)90242-H

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The gene (rsrIM) encoding the RsrI DNA methyltransferase (M . RsrI) from Rhodobacter sphaeroides was cloned and expressed in Escherichia coli. Under the control of a bacteriophage T7 promoter, 2% of the total protein in a crude extract was M . RsrI. This level of expression represents an approximately 50-fold increase over that present in the natural host. Chromatography using DNA cellulose and the S-adenosylmethionine analogue, sinefungin, was useful in purifying the enzyme to homogeneity. The purification yielded 100 times more enzyme than was obtained from the same quantity of R. sphaeroides cell paste. M.RsrI deposits one methyl group per productive DNA-binding event, as does its functional but sequence-nonhomologous analogue, M . EcoRI. Unlike M . EcoRI, the R. sphaeroides enzyme is a dimer at micromolar concentrations.

引用

页码：5 / 11

页数：7

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