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Experiment: Effect of the ribosomal protein mutation pgy1-1 on total mRNAs and on ribosome-bound mRNAs in Arabidopsis seedlings
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-656
Mutations in ribosomal proteins are known to cause specific developmental defects. For example, six different ribosomal protein mutations have been isolated as modifiers of the leaf patterning gene ASYMMETRIC LEAVES 1 (AS1). These modifiers, called piggyback (pgy), all condition ectopic outgrowths on the upper side of as1 mutant leaves (Pinon et al. 2008, Development 135, 1315).The specific developmental phenotypes of as1 pgy mutants suggested that ribosomes serve as a control point in expression of development genes. It is possible that different subpopulations of ribosomes, containing different ribosomal protein isoforms, are preferentially associated with translation or the translational control of subsets of mRNAs with specific roles in development.To test this hypothesis, we compared mRNAs pulled down with immunoprecipitated ribosomes in the pgy1-1 mutant and in the PGY1 control. To tag ribosomes for immunoprecipitation, we used pgy3-1 mutant plants complemented with a functional PGY3-FLAG fusion. As controls for changes in steady-state mRNA levels, we also compared RNA from total homogenates from pgy1-1 pgy3-1 PGY3-FLAG and from pgy3-1 PGY3-FLAG plants. Two independent experiments were performed, each using three biological replicates per treatment, comparing RNA extracted from total homogenates and from immunoprecipitated ribosomes (see detailed methods).
About the ExperimenterName: | Dr Cristina Pignocchi |
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Head of Lab Name: | Dr Robert Sablowski |
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Lab:
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Address: | Cell and Developmental Biology John Innes Centre Norwich Research Park
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Postcode:
| NR4 7UH |
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Country:
| United Kingdom |
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| Telephone Number:
| 441603450530 |
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All of the data available in this website/database is free, and you
are free to do whatever you please with it. If you intend to publish
work based on any of this data, please acknowledge us, contact the
experimenter above, and either acknowledge them or use them as
co-authors in the work.
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| About this ExperimentExperiment Type:
| gene_knock_out |
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Number of Slides: | 24 |
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| Experimental Parameters:
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parameter | gene_knock_out |
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Quality Control Measures Taken:
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References:
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Reference | Pinon et al. 2008, Development 135, 1315 |
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| Other Information:
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Slides in this Experiment
Hybridisation Set: Pignocchi: Effect of the ribosomal protein mutation pgy1-1 on total mRNAs and on ribosome-bound mRNAs in Arabidopsis seedlings
Slide: Pignocchi_exp2_wt_IP_rep1_ATH1 | | |
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Tissue:
| whole plant |
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Additional Organism Information:
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Sample Description | Generation of pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pignocchi_exp1_wt_TOT_rep2_ATH1 | | |
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Tissue:
| whole plant |
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Additional Organism Information:
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Sample Description | Generation of pgy3-1 PGY3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pignocchi_exp1_wt_IP_rep3_ATH1 | | |
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Tissue:
| whole plant |
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Additional Organism Information:
| |
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Sample Description | Generation of pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pignocchi_exp1_pgy1_TOT_rep2_ATH1 | | |
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Tissue:
| whole plant |
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Additional Organism Information:
| |
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Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
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|
Protocols for BioSource 1 |
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Slide: Pignocchi_exp1_pgy1_IP_rep1_ATH1 | | |
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Tissue:
| whole plant |
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Additional Organism Information:
| |
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Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Pignocchi_exp2_wt_TOT_rep3_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy3-1 PGY3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
|
|
Slide: Pignocchi_exp2_wt_TOT_rep2_ATH | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy3-1 PGY3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
|
|
Slide: Pignocchi_exp2_wt_TOT_rep1_ATH | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy3-1 PGY3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
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|
Slide: Pignocchi_exp2_wt_IP_rep3_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
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Sample Description | Generation of pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
|
|
Slide: Pignocchi_exp2_wt_IP_rep2_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Pignocchi_exp1_pgy1_IP_rep3_ATH1 | | |
|
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Tissue:
| whole plant |
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Additional Organism Information:
| |
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Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Pignocchi_exp1_pgy1_IP_rep2_ATH1 | | |
|
|
|
Tissue:
| whole plant |
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Additional Organism Information:
| |
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Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
|
|
Slide: Pignocchi_exp1_wt_IP_rep1_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
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|
Protocols for BioSource 1 |
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Slide: Pignocchi_exp1_pgy1_TOT_rep3_ATH1 | | |
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Tissue:
| whole plant |
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Additional Organism Information:
| |
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Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
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Slide: Pignocchi_exp1_pgy1_TOT_rep1_ATH1 | | |
|
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Tissue:
| whole plant |
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Additional Organism Information:
| |
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Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
|
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|
Slide: Pignocchi_exp1_wt_TOT_rep1_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy3-1 PGY3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
|
|
Slide: Pignocchi_exp1_wt_IP_rep2_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
|
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|
Slide: Pignocchi_exp2_pgy1_TOT_rep3_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
|
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|
Slide: Pignocchi_exp2_pgy1_TOT_rep2_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Pignocchi_exp2_pgy1_TOT_rep1_ATH1 | | |
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Tissue:
| whole plant |
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Additional Organism Information:
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Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
|
|
|
Slide: Pignocchi_exp2_pgy1_IP_rep3_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
---|
Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
|
|
Slide: Pignocchi_exp2_pgy1_IP_rep2_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
---|
Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
|
|
Slide: Pignocchi_exp2_pgy1_IP_rep1_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy1-1 pgy3-1 pgy3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy1-1 and pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then crossed and taken to F2 to produce homozygous PGY1 pgy3-1 PGY3p::PGY3-FLAG (wild-type control for PGY1) and pgy1-1 pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
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Other Information:
| |
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|
Protocols for BioSource 1 |
|
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|
Slide: Pignocchi_exp1_wt_TOT_rep3_ATH1 | | |
|
|
|
Tissue:
| whole plant |
---|
Additional Organism Information:
| |
---|
Sample Description | Generation of pgy3-1 PGY3-FLAG:
A 2.7 Kb genomic fragment encompassing At3g25520 (PGY3) was cloned into the binary vector pGWB10 to generate PGY3p::PGY3-FLAG. This construct was transformed into pgy3-1 (Pinon et al., 2008) using Agrobacterium mediated transformation (Clough and Bent, 1998). Transformants were selected based on complementation of pgy3-1 and on expression of PGY3-FLAG detected by western blot, then taken to F2 to produce pgy3-1 PGY3p::PGY3-FLAG.
Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
Pinon, V., Etchells, J.P., Rossignol, P., Collier, S.A., Arroyo, J.M., Martienssen, R.A., and Byrne, M.E. (2008). Three PIGGYBACK genes that specifically influence leaf patterning encode ribosomal proteins. Development 135, 1315-1324. |
---|
Other Information:
| |
---|
|
Protocols for BioSource 1 |
|
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|
Problems? Comments? Suggestions? Contact the Affymetrix Team