TRANSFECTION OF CHICKEN-EMBRYO FIBROBLASTS WITH MAREKS-DISEASE VIRUS-DNA

被引：86
作者：
MORGAN, RW
CANTELLO, JL
MCDERMOTT, CH
机构：
[1] Department of Animal Science and Agricultural Biochemistry, College of Agricultural Sciences, University of Delaware, Newark
来源：
AVIAN DISEASES | 1990年 / 34卷 / 02期
关键词：
D O I：
10.2307/1591417
中图分类号：
S85 [动物医学（兽医学）];
学科分类号：
0906 ;
摘要：
Total DNA from Marek's disease virus (MDV)-infected chicken embryo fibroblasts was transfected into freshly plated secondary chicken embryo fibroblasts using calcium phosphate-mediated transfection. Transfection frequencies were dose-dependent and non-linear. The maximum transfection frequencies of nine MDV DNA preparations using 8-25 micrograms total DNA ranged from 45 to 898 plaques per calcium phosphate/DNA precipitate. Approximately 100-200 plaques per 60-mm tissue-culture dish using 1-5 micrograms total DNA from MDV-infected chicken embryo fibroblasts were typically obtained. Transfection was most efficient when the pH of the HEPES buffer was 7.0, no additional carrier DNA was added to the precipitates, and the cultures were exposed for 3 minutes to 15% buffered glycerol 4 hours after the addition of the calcium phosphate/DNA precipitates.
引用
收藏
页码：345 / 351
页数：7
相关论文
共 39 条
1234