Transposon mutagenesis of baculoviruses: Analysis of Trichoplusia ni transposon IFP2 insertions within the FP-locus of nuclear polyhedrosis viruses

Lynne Csiszar Cary, Michael Goebel, Bartholomew G. Corsaro, Hwei Gene Wang, Elliot Rosen, M. J. Fraser

Research output: Contribution to journalArticle

342 Citations (Scopus)

Abstract

The transposable IFP2 element of Trichoplusia ni was originally isolated as a host DNA insertion in spontaneous FP mutants of Galleria mellonella or Autographa californica nuclear polyhedrosis viruses (NPVs). The termini of IFP2 insertions from five independently isolated FP mutants were sequenced. In all cases IFP2 is flanked by 13-bp terminal inverted repeats and has additional inverted repeats of 19 by in length located asymmetrically with respect to the ends of the element. Insertion of IFP2 into the viral genome always generated a duplication of the tetranucleotide target site, TTAA. There was an apparent preference for insertion within a 12-bp A+T-rich imperfect palindromic sequence surrounding the target site. Sequence analysis of three independent IFP2 elements revealed an internal domain of 2.475 kb containing an RNA polymerase II promoter region and two large open reading frames. Primer extension analysis of IFP2-specific mRNA positioned the 5′ terminus of the transcript. The element is present in DNA isolated from T. ni cell lines TN-368 and TN-5131, but is not apparent in DNAs isolated from the TN-R2 cell line or our laboratory colony of T. ni larvae, suggesting IFP2 was recently introduced into the T. ni genome.

Original languageEnglish (US)
Pages (from-to)156-169
Number of pages14
JournalVirology
Volume172
Issue number1
DOIs
StatePublished - 1989
Externally publishedYes

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Nucleopolyhedrovirus
Baculoviridae
Mutagenesis
Cell Line
DNA Transposable Elements
Terminal Repeat Sequences
RNA Polymerase II
Viral Genome
DNA
Genetic Promoter Regions
Open Reading Frames
Larva
Sequence Analysis
Genome
Messenger RNA
T-DNA

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Transposon mutagenesis of baculoviruses : Analysis of Trichoplusia ni transposon IFP2 insertions within the FP-locus of nuclear polyhedrosis viruses. / Cary, Lynne Csiszar; Goebel, Michael; Corsaro, Bartholomew G.; Wang, Hwei Gene; Rosen, Elliot; Fraser, M. J.

In: Virology, Vol. 172, No. 1, 1989, p. 156-169.

Research output: Contribution to journalArticle

Cary, Lynne Csiszar ; Goebel, Michael ; Corsaro, Bartholomew G. ; Wang, Hwei Gene ; Rosen, Elliot ; Fraser, M. J. / Transposon mutagenesis of baculoviruses : Analysis of Trichoplusia ni transposon IFP2 insertions within the FP-locus of nuclear polyhedrosis viruses. In: Virology. 1989 ; Vol. 172, No. 1. pp. 156-169.
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abstract = "The transposable IFP2 element of Trichoplusia ni was originally isolated as a host DNA insertion in spontaneous FP mutants of Galleria mellonella or Autographa californica nuclear polyhedrosis viruses (NPVs). The termini of IFP2 insertions from five independently isolated FP mutants were sequenced. In all cases IFP2 is flanked by 13-bp terminal inverted repeats and has additional inverted repeats of 19 by in length located asymmetrically with respect to the ends of the element. Insertion of IFP2 into the viral genome always generated a duplication of the tetranucleotide target site, TTAA. There was an apparent preference for insertion within a 12-bp A+T-rich imperfect palindromic sequence surrounding the target site. Sequence analysis of three independent IFP2 elements revealed an internal domain of 2.475 kb containing an RNA polymerase II promoter region and two large open reading frames. Primer extension analysis of IFP2-specific mRNA positioned the 5′ terminus of the transcript. The element is present in DNA isolated from T. ni cell lines TN-368 and TN-5131, but is not apparent in DNAs isolated from the TN-R2 cell line or our laboratory colony of T. ni larvae, suggesting IFP2 was recently introduced into the T. ni genome.",
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