Abstract
The split GFP technique is based on the auto-assembly of GFP when two polypeptides-GFP1-10 (residues 1-214; the detector) and GFP11 (residues 215-230; the tag)-both non-fluorescing on their own, associate spontaneously to form a fluorescent molecule. We evaluated this technique for its efficacy in contributing to the characterization of Cauliflower mosaic virus (CaMV) infection. A recombinant CaMV with GFP11 fused to the viral protein P6 (a key player in CaMV infection and major constituent of viral factory inclusions that arise during infection) was constructed and used to inoculate transgenic Arabidopsis thaliana expressing GFP1-10. The mutant virus (CaMV11P6) was infectious, aphid-transmissible and the insertion was stable over many passages. Symptoms on infected plants were delayed and milder. Viral protein accumulation, especially of recombinant 11P6, was greatly decreased, impeding its detection early in infection. Nonetheless, spread of infection from the inoculated leaf to other leaves was followed by whole plant imaging. Infected cells displayed in real time confocal laser scanning microscopy fluorescence in wild type-looking virus factories. Thus, it allowed for the first time to track a CaMV protein in vivo in the context of an authentic infection. 11P6 was immunoprecipitated with anti-GFP nanobodies, presenting a new application for the split GFP system in protein-protein interaction assays and proteomics. Taken together, split GFP can be an attractive alternative to using the entire GFP for protein tagging.
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Claims 7
Claim · model-written summary
The CaMV11P6 mutant virus was infectious, aphid-transmissible and its insertion stable over many passages.
- Taxon
- Arabidopsis thaliana
Arabidopsis thalianae1 - Interactor
- Cauliflower mosaic virus
Cauliflower mosaic viruse0pathogen - Outcome
- The mutant virus (CaMV11P6)e2
- Qualifier
- infectious, aphid-transmissible and the insertion was stable over many passagese2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“characterization of Cauliflower mosaic virus (CaMV) infection”
e1“transgenic Arabidopsis thaliana expressing GFP1-10”
e2“The mutant virus (CaMV11P6) was infectious, aphid-transmissible and the insertion was stable over many passages”
Claim · model-written summary
Symptoms on plants infected with recombinant CaMV11P6 were delayed and milder.
- Taxon
- Arabidopsis thalianaArabidopsis thalianae1
- Treatment
- A recombinant CaMV with GFP11 fused to the viral protein P6e2
- Interactor
- Cauliflower mosaic virusCauliflower mosaic viruse0pathogen
- Outcome
- Symptomse3
- Direction
- delayed and mildere3
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“characterization of Cauliflower mosaic virus (CaMV) infection”
e1“transgenic Arabidopsis thaliana expressing GFP1-10”
e3“Symptoms on infected plants were delayed and milder”
e2“A recombinant CaMV with GFP11 fused to the viral protein P6”
Claim · model-written summary
Viral protein accumulation of recombinant 11P6 was greatly decreased, impeding its early detection in infection.
- Taxon
- Arabidopsis thalianaArabidopsis thalianae1
- Material
- recombinant 11P6e4
- Treatment
- A recombinant CaMV with GFP11 fused to the viral protein P6e2
- Interactor
- Cauliflower mosaic virusCauliflower mosaic viruse0pathogen
- Outcome
- Viral protein accumulatione4
- Direction
- greatly decreasede4
- Qualifier
- impeding its detection early in infectione4
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“characterization of Cauliflower mosaic virus (CaMV) infection”
e1“transgenic Arabidopsis thaliana expressing GFP1-10”
e4“Viral protein accumulation, especially of recombinant 11P6, was greatly decreased, impeding its detection early in infection”
e2“A recombinant CaMV with GFP11 fused to the viral protein P6”
Claim · model-written summary
Infected cells showed fluorescence in wild type-looking virus factories in real-time confocal microscopy.
- Taxon
- Arabidopsis thalianaArabidopsis thalianae1
- Organ / stage
- Infected cellse5
- Outcome
- fluorescence in wild type-looking virus factoriese5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“transgenic Arabidopsis thaliana expressing GFP1-10”
e5“Infected cells displayed in real time confocal laser scanning microscopy fluorescence in wild type-looking virus factories”
Claim · model-written summary
Split GFP allowed tracking of a CaMV protein in vivo during an authentic infection.
- Taxon
- Arabidopsis thalianaArabidopsis thalianae1
- Interactor
- Cauliflower mosaic virusCauliflower mosaic viruse0pathogen
- Outcome
- track a CaMV protein in vivoe6
- Qualifier
- in the context of an authentic infectione6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“characterization of Cauliflower mosaic virus (CaMV) infection”
e1“transgenic Arabidopsis thaliana expressing GFP1-10”
e6“Thus, it allowed for the first time to track a CaMV protein in vivo in the context of an authentic infection”
Claim · model-written summary
The 11P6 protein was immunoprecipitated with anti-GFP nanobodies.
- Taxon
- Arabidopsis thalianaArabidopsis thalianae1
- Gene
- 11P6e7
- Outcome
- immunoprecipitated with anti-GFP nanobodiese7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“transgenic Arabidopsis thaliana expressing GFP1-10”
e7“11P6 was immunoprecipitated with anti-GFP nanobodies, presenting a new application for the split GFP system in protein-protein interaction assays and proteomics”
Claim · model-written summary
Split GFP is an attractive alternative to entire GFP for protein tagging.
Verbatim
e0“split GFP can be an attractive alternative to using the entire GFP for protein tagging”
The same questions in other papers
Brassica oleracea var. botrytis · Cauliflower mosaic virus · Arabidopsis thaliana (thale cress)Positive and negative results
1 in the opposite direction
- negative
Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.