Bromodeoxyuridine-labeled viral particles as a tool for visualization of the immediate-early events of human cytomegalovirus infection

被引:25
作者
Rosenke, K [1 ]
Fortunato, EA [1 ]
机构
[1] Univ Idaho, Dept Microbiol Mol Biol & Biochem, Ctr Reprod Biol, Moscow, ID 83844 USA
关键词
D O I
10.1128/JVI.78.14.7818-7822.2004
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We describe here a simple method for labeling the genome of human cytomegalovirus, a large double-stranded DNA virus, with bromodeoxyuridine (BrdU). The labeled DNA was incorporated into viral particles, which were then collected in cell supernatant. To demonstrate the versatility and effectiveness of this method, labeled virions were used to study the immediate-early events of virus-host cell interaction via indirect immunofluorescence microscopy. It is our hope that this new methodology will prove useful in the study of binding, entry and viral genome deposition in diverse virus systems.
引用
收藏
页码:7818 / 7822
页数:5
相关论文
共 23 条
[1]  
ALBERTS B, 1994, MOL BIOL CELL, P1207
[2]   Requirement for p53 and p21 to sustain G2 arrest after DNA damage [J].
Bunz, F ;
Dutriaux, A ;
Lengauer, C ;
Waldman, T ;
Zhou, S ;
Brown, JP ;
Sedivy, JM ;
Kinzler, KW ;
Vogelstein, B .
SCIENCE, 1998, 282 (5393) :1497-1501
[3]   Structure of the human cytomegalovirus B capsid by electron cryomicroscopy and image reconstruction [J].
Butcher, SJ ;
Aitken, J ;
Mitchell, J ;
Gowen, B ;
Dargan, DJ .
JOURNAL OF STRUCTURAL BIOLOGY, 1998, 124 (01) :70-76
[4]   HUMAN CYTOMEGALOVIRUS PENETRATES HOST-CELLS BY PH-INDEPENDENT FUSION AT THE CELL-SURFACE [J].
COMPTON, T ;
NEPOMUCENO, RR ;
NOWLIN, DM .
VIROLOGY, 1992, 191 (01) :387-395
[5]   Incorporation of the green fluorescent protein into the herpes simplex virus type 1 capsid [J].
Desai, P ;
Person, S .
JOURNAL OF VIROLOGY, 1998, 72 (09) :7563-7568
[6]   Live-cell analysis of a green fluorescent protein-tagged herpes simplex virus infection [J].
Elliott, G ;
O'Hare, P .
JOURNAL OF VIROLOGY, 1999, 73 (05) :4110-4119
[7]   Directional spread of an α-herpesvirus in the nervous system [J].
Enquist, LW ;
Tomishima, MJ ;
Gross, S ;
Smith, GA .
VETERINARY MICROBIOLOGY, 2002, 86 (1-2) :5-16
[8]   Specific chromosome 1 breaks induced by human cytomegalovirus [J].
Fortunato, EA ;
Dell'Aquila, ML ;
Spector, DH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (02) :853-858
[9]   p53 and RPA are sequestered in viral replication centers in the nuclei of cells infected with human cytomegalovirus [J].
Fortunato, EA ;
Spector, DH .
JOURNAL OF VIROLOGY, 1998, 72 (03) :2033-2039
[10]   Human cytomegalovirus immediate early interaction with host nuclear structures: Definition of an immediate transcript environment [J].
Ishov, AM ;
Stenberg, RM ;
Maul, GG .
JOURNAL OF CELL BIOLOGY, 1997, 138 (01) :5-16