Growth suppression by members of the retinoblastoma protein family.
Growth suppression by members of the retinoblastoma protein family.
复制标题
视网膜母细胞瘤蛋白家族成员的生长抑制。
DOI：
10.1101/sqb.1994.059.01.011
复制
发表时间：
1994
期刊：
影响因子：
--
通讯作者：
E. Harlow
中科院分区：
文献类型：
--
作者：
L. Zhu;G. Enders;C. L. Wu;M. Starz;K. Moberg;J. Lees;N. Dyson;E. Harlow
图1. pRB和p107是E2 F活性的阻遏物。使用磷酸钙沉淀法用以下质粒瞬时转染Saos-2细胞：5 yg CAT报告质粒（如所示的E2-CAT、E2-CAT [E2 F-]或E2-CAT [ATF-]）; 5 yg pRSVfl-gal; 5 yg CMV表达构建体（如所示的载体pCMVpRB或pCMVp 107）和5 yg pCMVE 1A 12 S。用pBSK DNA调节转染的DNA总量为25 kg。转染后40小时制备细胞提取物。3-用O-硝基苯酚-β-D-半乳糖苷法测定gal活性，并以实际OD 420阅读表示。将CAT活性相对于fl-gal活性标准化，以任意单位表示。
Figure 1. pRB and p107 are repressors of E2F activity. Saos-2 cells were transiently transfected using the calcium phosphate precipitation method with the following plasmids: 5/~ g CAT reporter plasmid (E2-CAT, E2-CAT [E2F-], or E2-CAT [ATF-] as indicated); 5~ g pRSVfl-gal; 5/~ g of the CMV expression constructs (vector, pCMVpRB, or pCMVpl07 as indicated) and 5/~ g of pCMVE1A12S as indicated. The total amounts of DNA in transfection were adjusted to 25 k~ g with pBSK DNA. Cell extracts were prepared 40 hr after transfection./3-gal activity was determined with the O-nitrophenol-fl-D-galactoside method and presented as the actual OD420 reading. CAT activity was normalized against the fl-gal activity to be presented as arbitrary units.