LOCALIZATION OF ADENOVIRUS DNA BY INSITU HYBRIDIZATION ELECTRON-MICROSCOPY

被引:10
作者
JIAO, RJ [1 ]
YU, WD [1 ]
DING, MX [1 ]
ZHAI, ZH [1 ]
机构
[1] BEIJING UNIV, DEPT BIOL, BEIJING 100871, PEOPLES R CHINA
关键词
AUTORADIOGRAPHY; NICK TRANSLATION; NUCLEAR MATRIX; BIOTINATION; DNA REPLICATION;
D O I
10.1002/jemt.1070210104
中图分类号
R602 [外科病理学、解剖学]; R32 [人体形态学];
学科分类号
100101 ;
摘要
Biotinylated deoxyadenosine triphosphate (dATP) (Bio-7-dATP) and H-3 deoxythymidine triphosphate (dTTP) labeled adenovirus DNA were hybridized in situ to thin sections of Lowicryl K4M-embedded and whole-mount extracted HeLa cells infected with adenovirus. The biotinylated probe was detected by exposing the extracted cells or sections to antibodies against biotin followed by colloidal gold-conjugated secondary antibodies and then critical-point dried while H-3-dTTP labeled probe by electron microscopic autoradiography. On Lowicryl K4M sections, gold particles and silver grains were mainly restricted in the nucleus. Furthermore, whole-mount results suggested that replicating adenovirus DNA is localized on the nuclear matrix of its host cell. In this paper, the described non-radioactive procedures for hybrid detection offered several advantages: a) rapid signal detection; b) superior morphological preservation and spatial resolution; c) precise localization; and d) on Lowicryl K4M sections, signal to noise equivalent to radiolabeling.
引用
收藏
页码:23 / 31
页数:9
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