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Deletion analysis of a replication origin of human cytomegalovirus by a novel assay system with a combination of microinjection and polymerase chain reaction.

Abstract：

:We have developed a sensitive assay for DNA replication in mammalian cells that enables us to detect replicated DNA fragments of human cytomegalovirus. A 1/100 portion of DNA extracted from 1000 microinjected cells was subjected to polymerase chain reaction after digestion with appropriate restriction enzymes to differentiate replicated from nonreplicated plasmids. A portion of the amplified DNA was electrophoresed to detect replicated DNA. Subfragments of the HindIII fragment A (24 kb, map units 0.37-0.47) of strain Towne, which contains a previously identified origin of DNA replication (oriLyt), were analyzed by the assay system. A 4.3-kb subfragment (AatII-SacI) replicated as efficiently as the HindIII fragment A. Efficient replication ability was lost with a 1.3-kb deletion from the AatII end or a 0.9-kb deletion from the SacI end. These results suggest that the boundaries of oriLyt of Human cytomegalovirus strain Towne lie within the 1.3- and the 0.9-kb regions of the 4.3-kb fragment.

journal_name

Virology

journal_title

Virology

authors

Watanabe S,Yamaguchi N

doi

10.1006/viro.1993.1038

subject

Has Abstract

pub_date

1993-01-01 00:00:00

pages

332-5

issue

1

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(83)71038-X

journal_volume

192

pub_type

杂志文章

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