PMID- 25835531
OWN - NLM
STAT- MEDLINE
DCOM- 20151217
LR  - 20191210
IS  - 1999-4915 (Electronic)
IS  - 1999-4915 (Linking)
VI  - 7
IP  - 4
DP  - 2015 Mar 31
TI  - Development of a recombination system for the generation of occlusion positive 
      genetically modified Anticarsia gemmatalis multiple nucleopolyhedrovirus.
PG  - 1599-612
LID - 10.3390/v7041599 [doi]
AB  - Anticarsia gemmatalis is an important pest in legume crops in South America and 
      it has been successfully controlled using Anticarsia gemmatalis Multiple 
      Nucleopolyhedrovirus (AgMNPV) in subtropical climate zones. Nevertheless, in 
      temperate climates its speed of kill is too slow. Taking this into account, 
      genetic modification of AgMNPV could lead to improvements of its biopesticidal 
      properties. Here we report the generation of a two-component system that allows 
      the production of recombinant AgMNPV. This system is based on a parental AgMNPV 
      in which the polyhedrin gene (polh) was replaced by a bacterial beta-galactosidase 
      (lacZ) gene flanked by two target sites for the homing endonuclease I-PpoI. 
      Co-transfection of insect cells with linearized (I-PpoI-digested) parental genome 
      and a transfer vector allowed the restitution of polh and the expression of a 
      heterologous gene upon homologous recombination, with a low background of 
      non-recombinant AgMNPV. The system was validated by constructing a recombinant 
      occlusion-positive (polh+) AgMNPV expressing the green fluorescent protein gene 
      (gfp). This recombinant virus infected larvae normally per os and led to the 
      expression of GFP in cell culture as well as in A. gemmatalis larvae. These 
      results demonstrate that the system is an efficient method for the generation of 
      recombinant AgMNPV expressing heterologous genes, which can be used for manifold 
      purposes, including biotechnological and pharmaceutical applications and the 
      production of orally infectious recombinants with improved biopesticidal 
      properties.
FAU - Haase, Santiago
AU  - Haase S
AD  - Instituto de Biotecnologia y Biologia Molecular (IBBM), Departamento de Ciencias 
      Biologicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 
      CONICET, 1900-La Plata, Argentina. shaase@biol.unlp.edu.ar.
FAU - McCarthy, Christina B
AU  - McCarthy CB
AD  - Instituto de Biotecnologia y Biologia Molecular (IBBM), Departamento de Ciencias 
      Biologicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 
      CONICET, 1900-La Plata, Argentina. cmccarthy@conicet.gov.ar.
AD  - Centro Regional de Estudios Genomicos (CREG), Facultad de Ciencias Exactas, 
      Universidad Nacional de La Plata, 1900-La Platae, Argentina. 
      cmccarthy@conicet.gov.ar.
FAU - Ferrelli, M Leticia
AU  - Ferrelli ML
AD  - Instituto de Biotecnologia y Biologia Molecular (IBBM), Departamento de Ciencias 
      Biologicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 
      CONICET, 1900-La Plata, Argentina. lferrelli@biol.unlp.edu.ar.
FAU - Pidre, Matias L
AU  - Pidre ML
AD  - Instituto de Biotecnologia y Biologia Molecular (IBBM), Departamento de Ciencias 
      Biologicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 
      CONICET, 1900-La Plata, Argentina. mlpidre@biol.unlp.edu.ar.
FAU - Sciocco-Cap, Alicia
AU  - Sciocco-Cap A
AD  - Instituto de Microbiologia y Zoologia Agricola (IMYZA), Instituto Nacional de 
      Tecnologia Agropecuaria (INTA), 1712-Castelar, Argentina. 
      sciocco.alicia@inta.gob.ar.
FAU - Romanowski, Victor
AU  - Romanowski V
AD  - Instituto de Biotecnologia y Biologia Molecular (IBBM), Departamento de Ciencias 
      Biologicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 
      CONICET, 1900-La Plata, Argentina. victor@biol.unlp.edu.ar.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150331
PL  - Switzerland
TA  - Viruses
JT  - Viruses
JID - 101509722
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Entomology/*methods
MH  - Genes, Reporter
MH  - Green Fluorescent Proteins/analysis/genetics
MH  - Larva/virology
MH  - Lepidoptera/*virology
MH  - Molecular Biology/*methods
MH  - Nucleopolyhedroviruses/*genetics/*growth & development
MH  - *Recombination, Genetic
MH  - South America
MH  - Virology/*methods
PMC - PMC4411667
EDAT- 2015/04/04 06:00
MHDA- 2015/12/19 06:00
PMCR- 2015/04/01
CRDT- 2015/04/04 06:00
PHST- 2015/01/14 00:00 [received]
PHST- 2015/03/12 00:00 [revised]
PHST- 2015/03/26 00:00 [accepted]
PHST- 2015/04/04 06:00 [entrez]
PHST- 2015/04/04 06:00 [pubmed]
PHST- 2015/12/19 06:00 [medline]
PHST- 2015/04/01 00:00 [pmc-release]
AID - v7041599 [pii]
AID - viruses-07-01599 [pii]
AID - 10.3390/v7041599 [doi]
PST - epublish
SO  - Viruses. 2015 Mar 31;7(4):1599-612. doi: 10.3390/v7041599.