| Purification and Characterization of DNA Methylase From HL-60 Cells |
| |
| 引用本文： | 何忠效,王顺泰,姚知行,王秀奇. Purification and Characterization of DNA Methylase From HL-60 Cells[J]. 中国科学B辑(英文版), 1994, 0(4) |
| |
| 作者姓名： | 何忠效 王顺泰 姚知行 王秀奇 |
| |
| 作者单位： | Department of Biology，Beijing Normal University，Beijing 100875，PRC，State Key Laboratory of Macrobiomolecule，Beijing 100101，PRC |
| |
| 基金项目： | Project supported by the National Natural Science Foundation of China. |
| |
| 摘 要： | A solid leukemia sarcoma has been successfully developed after subcutaneous inoculation of the cultured human promyelocytic leukemia cells (HL-60 cells) into unde mice. The solid leukemia sarcoma is a more plantiful source than the cultured cells for enzymatic study and its growing environment is closer to that of the human body than the cultured cells.We establish an efficient procedure of purifying HL-60 cells DNA methylase which includes: disruption of HL-60 cells by homogenization and sonication, removing the cell fragments and cellular particles by centrifuge and ultracentrifuge (105.000 g); removing endogenous DNA by streptomycin sulfate, salting out by (NH4)2SO4, ion exchange chromatography on DEAE-cellulose (DE-52), gel filtration over Sephadex G-100 column.The DNA methylase from HL-60 cells has been purified 204 fold by this procedure. The purified enzyme shows a single-band on PG-PAGE. A 479-kD molecular weight of this enzyme is measured by PG-PAGE. The enzyme properties of HL-60 DNA methylase
|
Purification and Characterization of DNA Methylase From HL-60 Cells |
| |
| HE Zhong-Xiao,WANG Shun-TaiYAO Zhi-Xing and WANG Xiu-Qi. Purification and Characterization of DNA Methylase From HL-60 Cells[J]. Science in China(Chemistry), 1994, 0(4) |
| |
| Authors: | HE Zhong-Xiao WANG Shun-TaiYAO Zhi-Xing WANG Xiu-Qi |
| |
| Abstract: | A solid leukemia sarcoma has been successfully developed after subcutaneous inoculation of the cultured human promyelocytic leukemia cells (HL-60 cells) into unde mice. The solid leukemia sarcoma is a more plantiful source than the cultured cells for enzymatic study and its growing environment is closer to that of the human body than the cultured cells.We establish an efficient procedure of purifying HL-60 cells DNA methylase which includes: disruption of HL-60 cells by homogenization and sonication, removing the cell fragments and cellular particles by centrifuge and ultracentrifuge (105.000 g); removing endogenous DNA by streptomycin sulfate, salting out by (NH4)2SO4, ion exchange chromatography on DEAE-cellulose (DE-52), gel filtration over Sephadex G-100 column.The DNA methylase from HL-60 cells has been purified 204 fold by this procedure. The purified enzyme shows a single-band on PG-PAGE. A 479-kD molecular weight of this enzyme is measured by PG-PAGE. The enzyme properties of HL-60 DNA methylase are also studied. |
| |
| Keywords: | HL-60 cells DNA methylase chromatography PG-PAGE. |
| 本文献已被 CNKI 等数据库收录！ |
