Functional analysis of FP25K of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus

D. Wu, F. Deng, X. Sun, H. Wang, L. Yuan, J.M. Vlak, Z.H. Hu

Research output: Contribution to journalArticleAcademicpeer-review

25 Citations (Scopus)

Abstract

The fp25k gene of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) was studied. HearNPV fp25k gene transcription was found starting from about 18 h post-infection, and protein could be detected from the same time with antiserum against FP25K. To study the function of HearNPV fp25k, a recombinant HearNPV (HaBacWD11) with an enhanced green fluorescent protein (GFP) gene replacing the fp25k was constructed using HaBacHZ8, a bacmid of HearNPV that lacks the polyhedrin gene. Growth curve analysis showed that HaBacWD11 produced higher titres of budded viruses (BVs) than its wild-type counterpart HaBacHZ8¿GFP. Electron microscopic analysis indicated that at the late stage of infection, the number of intranuclear enveloped nucleocapsids in HaBacWD11-infected cells was much less than that of HaBacHZ8¿GFP. A rescue recombinant virus HaBacWD14 was constructed by reintroducing fp25k gene into HaBacWD11. The growth curve and electron microscopic analysis of the rescued recombinant confirmed that the increase of BV yield and the decrease of the virion production in infected cells were the result of fp25k deletion. The expression of membrane fusion protein (Ha133) and ODV-E66 were studied using the FP25K mutants HaBacWD11 and HaBacHZ8¿GFP. Unlike FP25K mutants in Autographa californica multicapsid NPV (AcMNPV), which caused an increase in the expression of membrane fusion protein GP64 and a decrease of ODV-E66, no obvious changes at the expression level of Ha133 and ODV-E66 were observed in HearNPV FP25K mutant.
Original languageEnglish
Pages (from-to)2439-2444
JournalJournal of General Virology
Volume86
Issue number9
DOIs
Publication statusPublished - 2005

Fingerprint

Nucleopolyhedrovirus
Nucleocapsid
Membrane Fusion Proteins
Genes
Electrons
Viruses
Growth
Infection
Viral Load
Virion
Immune Sera

Keywords

  • nuclear polyhedrosis-virus
  • lymantria-dispar nucleopolyhedrovirus
  • 25k fp gene
  • autographa-californica
  • serial passage
  • baculovirus
  • mutations
  • protein
  • identification
  • transport

Cite this

Wu, D. ; Deng, F. ; Sun, X. ; Wang, H. ; Yuan, L. ; Vlak, J.M. ; Hu, Z.H. / Functional analysis of FP25K of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus. In: Journal of General Virology. 2005 ; Vol. 86, No. 9. pp. 2439-2444.
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title = "Functional analysis of FP25K of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus",
abstract = "The fp25k gene of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) was studied. HearNPV fp25k gene transcription was found starting from about 18 h post-infection, and protein could be detected from the same time with antiserum against FP25K. To study the function of HearNPV fp25k, a recombinant HearNPV (HaBacWD11) with an enhanced green fluorescent protein (GFP) gene replacing the fp25k was constructed using HaBacHZ8, a bacmid of HearNPV that lacks the polyhedrin gene. Growth curve analysis showed that HaBacWD11 produced higher titres of budded viruses (BVs) than its wild-type counterpart HaBacHZ8¿GFP. Electron microscopic analysis indicated that at the late stage of infection, the number of intranuclear enveloped nucleocapsids in HaBacWD11-infected cells was much less than that of HaBacHZ8¿GFP. A rescue recombinant virus HaBacWD14 was constructed by reintroducing fp25k gene into HaBacWD11. The growth curve and electron microscopic analysis of the rescued recombinant confirmed that the increase of BV yield and the decrease of the virion production in infected cells were the result of fp25k deletion. The expression of membrane fusion protein (Ha133) and ODV-E66 were studied using the FP25K mutants HaBacWD11 and HaBacHZ8¿GFP. Unlike FP25K mutants in Autographa californica multicapsid NPV (AcMNPV), which caused an increase in the expression of membrane fusion protein GP64 and a decrease of ODV-E66, no obvious changes at the expression level of Ha133 and ODV-E66 were observed in HearNPV FP25K mutant.",
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author = "D. Wu and F. Deng and X. Sun and H. Wang and L. Yuan and J.M. Vlak and Z.H. Hu",
year = "2005",
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Functional analysis of FP25K of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus. / Wu, D.; Deng, F.; Sun, X.; Wang, H.; Yuan, L.; Vlak, J.M.; Hu, Z.H.

In: Journal of General Virology, Vol. 86, No. 9, 2005, p. 2439-2444.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Functional analysis of FP25K of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus

AU - Wu, D.

AU - Deng, F.

AU - Sun, X.

AU - Wang, H.

AU - Yuan, L.

AU - Vlak, J.M.

AU - Hu, Z.H.

PY - 2005

Y1 - 2005

N2 - The fp25k gene of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) was studied. HearNPV fp25k gene transcription was found starting from about 18 h post-infection, and protein could be detected from the same time with antiserum against FP25K. To study the function of HearNPV fp25k, a recombinant HearNPV (HaBacWD11) with an enhanced green fluorescent protein (GFP) gene replacing the fp25k was constructed using HaBacHZ8, a bacmid of HearNPV that lacks the polyhedrin gene. Growth curve analysis showed that HaBacWD11 produced higher titres of budded viruses (BVs) than its wild-type counterpart HaBacHZ8¿GFP. Electron microscopic analysis indicated that at the late stage of infection, the number of intranuclear enveloped nucleocapsids in HaBacWD11-infected cells was much less than that of HaBacHZ8¿GFP. A rescue recombinant virus HaBacWD14 was constructed by reintroducing fp25k gene into HaBacWD11. The growth curve and electron microscopic analysis of the rescued recombinant confirmed that the increase of BV yield and the decrease of the virion production in infected cells were the result of fp25k deletion. The expression of membrane fusion protein (Ha133) and ODV-E66 were studied using the FP25K mutants HaBacWD11 and HaBacHZ8¿GFP. Unlike FP25K mutants in Autographa californica multicapsid NPV (AcMNPV), which caused an increase in the expression of membrane fusion protein GP64 and a decrease of ODV-E66, no obvious changes at the expression level of Ha133 and ODV-E66 were observed in HearNPV FP25K mutant.

AB - The fp25k gene of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) was studied. HearNPV fp25k gene transcription was found starting from about 18 h post-infection, and protein could be detected from the same time with antiserum against FP25K. To study the function of HearNPV fp25k, a recombinant HearNPV (HaBacWD11) with an enhanced green fluorescent protein (GFP) gene replacing the fp25k was constructed using HaBacHZ8, a bacmid of HearNPV that lacks the polyhedrin gene. Growth curve analysis showed that HaBacWD11 produced higher titres of budded viruses (BVs) than its wild-type counterpart HaBacHZ8¿GFP. Electron microscopic analysis indicated that at the late stage of infection, the number of intranuclear enveloped nucleocapsids in HaBacWD11-infected cells was much less than that of HaBacHZ8¿GFP. A rescue recombinant virus HaBacWD14 was constructed by reintroducing fp25k gene into HaBacWD11. The growth curve and electron microscopic analysis of the rescued recombinant confirmed that the increase of BV yield and the decrease of the virion production in infected cells were the result of fp25k deletion. The expression of membrane fusion protein (Ha133) and ODV-E66 were studied using the FP25K mutants HaBacWD11 and HaBacHZ8¿GFP. Unlike FP25K mutants in Autographa californica multicapsid NPV (AcMNPV), which caused an increase in the expression of membrane fusion protein GP64 and a decrease of ODV-E66, no obvious changes at the expression level of Ha133 and ODV-E66 were observed in HearNPV FP25K mutant.

KW - nuclear polyhedrosis-virus

KW - lymantria-dispar nucleopolyhedrovirus

KW - 25k fp gene

KW - autographa-californica

KW - serial passage

KW - baculovirus

KW - mutations

KW - protein

KW - identification

KW - transport

U2 - 10.1099/vir.0.81110-0

DO - 10.1099/vir.0.81110-0

M3 - Article

VL - 86

SP - 2439

EP - 2444

JO - Journal of General Virology

JF - Journal of General Virology

SN - 0022-1317

IS - 9

ER -