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Selection of Marek's disease virus recombinants expressing the Escherichia coli gpt gene.

Abstract：

:We developed a positive selection method for recovering Marek's disease virus (MDV) recombinants. The Escherichia coli xanthine-guanine phosphoribosyltransferase gene (gpt), under the control of the major immediate-early promoter from cytomegalovirus, was inserted into the inverted repeats flanking the unique long (UL) region of a non-pathogenic serotype 2 MDV strain 281MI/1. In a second demonstration of the usefulness of the positive selection system, the gpt gene was inserted into the inverted repeats flanking the unique short (US) region of the turkey herpesvirus (HVT) strain FC126. The targeted insertion site in 281MI/1 was in a previously established nonessential site for virus replication. The targeted insertion site for FC126, at the junction of the UL and US regions, is a nonessential site for in vitro replication of herpes simplex virus. Recombinant viruses were easily selected by incubating the transfected cells in mycophenolic acid (MPA)-containing medium. Purification of recombinants resulted from a series of trypsinization and sonication steps combined with the culturing of virus in MPA-containing medium to inhibit wild-type virus replication. This simple technique for recovering MDV and HVT recombinants should increase the efficiency of identifying nonessential sites and gene function analysis by insertional mutagenesis.

journal_name

Virology

journal_title

Virology

authors

Marshall DR,Reilly JD,Liu X,Silva RF

doi

10.1006/viro.1993.1415

subject

Has Abstract

pub_date

1993-08-01 00:00:00

pages

638-48

issue

2

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(83)71415-7

journal_volume

195

pub_type

杂志文章

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