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TaKaRa puasp svb
Effect of modified forms of the <t>Svb</t> protein on epidermal trichome formation. <t>UAS-GFP</t> (control) ( A,A’ ), UAS-Svb-ACT ( B,B’ ) and UAS-Svb-3Kmut ( C,C’ ) were expressed in the embryonic epidermis under the control of the ptc-Gal4 driver. Top rows show whole embryo cuticles ( A–C ), the bottom row shows close-ups in the ventral region of the third abdominal segment ( A’–C’ ). Svb-ACT, which lacks the N-terminal repressor domain and thus mimics the processed form of Svb, acts as a constitutive activator of transcription and triggers the production of ectopic trichomes. In contrast, Svb-3Kmut -bearing mutations on the 3 Lysines ubiquitinated by Ubr3 in response to Tal peptides- behaves as a repressor and counteracts endogenous Svb activity, resulting in loss of trichomes.
Puasp Svb, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/puasp svb/product/TaKaRa
Average 93 stars, based on 2 article reviews
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puasp svb - by Bioz Stars, 2020-08
93/100 stars

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1) Product Images from "The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning"

Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning

Journal: eLife

doi: 10.7554/eLife.39748

Effect of modified forms of the Svb protein on epidermal trichome formation. UAS-GFP (control) ( A,A’ ), UAS-Svb-ACT ( B,B’ ) and UAS-Svb-3Kmut ( C,C’ ) were expressed in the embryonic epidermis under the control of the ptc-Gal4 driver. Top rows show whole embryo cuticles ( A–C ), the bottom row shows close-ups in the ventral region of the third abdominal segment ( A’–C’ ). Svb-ACT, which lacks the N-terminal repressor domain and thus mimics the processed form of Svb, acts as a constitutive activator of transcription and triggers the production of ectopic trichomes. In contrast, Svb-3Kmut -bearing mutations on the 3 Lysines ubiquitinated by Ubr3 in response to Tal peptides- behaves as a repressor and counteracts endogenous Svb activity, resulting in loss of trichomes.
Figure Legend Snippet: Effect of modified forms of the Svb protein on epidermal trichome formation. UAS-GFP (control) ( A,A’ ), UAS-Svb-ACT ( B,B’ ) and UAS-Svb-3Kmut ( C,C’ ) were expressed in the embryonic epidermis under the control of the ptc-Gal4 driver. Top rows show whole embryo cuticles ( A–C ), the bottom row shows close-ups in the ventral region of the third abdominal segment ( A’–C’ ). Svb-ACT, which lacks the N-terminal repressor domain and thus mimics the processed form of Svb, acts as a constitutive activator of transcription and triggers the production of ectopic trichomes. In contrast, Svb-3Kmut -bearing mutations on the 3 Lysines ubiquitinated by Ubr3 in response to Tal peptides- behaves as a repressor and counteracts endogenous Svb activity, resulting in loss of trichomes.

Techniques Used: Modification, Activated Clotting Time Assay, Activity Assay

Schematic representation of Tc-shavenbaby locus ( A ) and transcript ( B ), showing the site at which a GFP-containing marker plasmid was inserted by CRISPR/cas9 genome editing (see also Materials and methods). The mutagenic cassette is inserted into exon 2, that is within the open reading frame upstream of the region encoding the DNA-binding zinc finger domain. Gene disruption leads to mRNA truncation after the insertion site, since Tc-svb expression is absent in homozygous mutants. In addition to segmentation defects and transformation toward thoracic identity, other phenotypes observed in Tc-svb CRISPR mutants include incipient spiracles (possibly a secondary effect of cuticle thinning leading to a defect in the development of tracheal rings); sensory bristles that are shorter and thicker; leg segment boundaries that are not clearly defined; missing leg bristles; unsclerotized pretarsi with soft, rounded apices; and antennae lacking the terminal setae. Therefore, late functions of Tc-svb in epidermal and appendage differentiation are strongly affected in Tc-svb CRISPR mutant embryos, while the segmentation phenotype is milder that Tc-svb -RNAi knockdown due to maternal contribution of Tc-svb (Ray et al., in preparation).
Figure Legend Snippet: Schematic representation of Tc-shavenbaby locus ( A ) and transcript ( B ), showing the site at which a GFP-containing marker plasmid was inserted by CRISPR/cas9 genome editing (see also Materials and methods). The mutagenic cassette is inserted into exon 2, that is within the open reading frame upstream of the region encoding the DNA-binding zinc finger domain. Gene disruption leads to mRNA truncation after the insertion site, since Tc-svb expression is absent in homozygous mutants. In addition to segmentation defects and transformation toward thoracic identity, other phenotypes observed in Tc-svb CRISPR mutants include incipient spiracles (possibly a secondary effect of cuticle thinning leading to a defect in the development of tracheal rings); sensory bristles that are shorter and thicker; leg segment boundaries that are not clearly defined; missing leg bristles; unsclerotized pretarsi with soft, rounded apices; and antennae lacking the terminal setae. Therefore, late functions of Tc-svb in epidermal and appendage differentiation are strongly affected in Tc-svb CRISPR mutant embryos, while the segmentation phenotype is milder that Tc-svb -RNAi knockdown due to maternal contribution of Tc-svb (Ray et al., in preparation).

Techniques Used: Marker, Plasmid Preparation, CRISPR, Binding Assay, Expressing, Transformation Assay, Mutagenesis

2) Product Images from "The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning"

Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning

Journal: eLife

doi: 10.7554/eLife.39748

Effect of modified forms of the Svb protein on epidermal trichome formation. UAS-GFP (control) ( A,A’ ), UAS-Svb-ACT ( B,B’ ) and UAS-Svb-3Kmut ( C,C’ ) were expressed in the embryonic epidermis under the control of the ptc-Gal4 driver. Top rows show whole embryo cuticles ( A–C ), the bottom row shows close-ups in the ventral region of the third abdominal segment ( A’–C’ ). Svb-ACT, which lacks the N-terminal repressor domain and thus mimics the processed form of Svb, acts as a constitutive activator of transcription and triggers the production of ectopic trichomes. In contrast, Svb-3Kmut -bearing mutations on the 3 Lysines ubiquitinated by Ubr3 in response to Tal peptides- behaves as a repressor and counteracts endogenous Svb activity, resulting in loss of trichomes.
Figure Legend Snippet: Effect of modified forms of the Svb protein on epidermal trichome formation. UAS-GFP (control) ( A,A’ ), UAS-Svb-ACT ( B,B’ ) and UAS-Svb-3Kmut ( C,C’ ) were expressed in the embryonic epidermis under the control of the ptc-Gal4 driver. Top rows show whole embryo cuticles ( A–C ), the bottom row shows close-ups in the ventral region of the third abdominal segment ( A’–C’ ). Svb-ACT, which lacks the N-terminal repressor domain and thus mimics the processed form of Svb, acts as a constitutive activator of transcription and triggers the production of ectopic trichomes. In contrast, Svb-3Kmut -bearing mutations on the 3 Lysines ubiquitinated by Ubr3 in response to Tal peptides- behaves as a repressor and counteracts endogenous Svb activity, resulting in loss of trichomes.

Techniques Used: Modification, Activated Clotting Time Assay, Activity Assay

Schematic representation of Tc-shavenbaby locus ( A ) and transcript ( B ), showing the site at which a GFP-containing marker plasmid was inserted by CRISPR/cas9 genome editing (see also Materials and methods). The mutagenic cassette is inserted into exon 2, that is within the open reading frame upstream of the region encoding the DNA-binding zinc finger domain. Gene disruption leads to mRNA truncation after the insertion site, since Tc-svb expression is absent in homozygous mutants. In addition to segmentation defects and transformation toward thoracic identity, other phenotypes observed in Tc-svb CRISPR mutants include incipient spiracles (possibly a secondary effect of cuticle thinning leading to a defect in the development of tracheal rings); sensory bristles that are shorter and thicker; leg segment boundaries that are not clearly defined; missing leg bristles; unsclerotized pretarsi with soft, rounded apices; and antennae lacking the terminal setae. Therefore, late functions of Tc-svb in epidermal and appendage differentiation are strongly affected in Tc-svb CRISPR mutant embryos, while the segmentation phenotype is milder that Tc-svb -RNAi knockdown due to maternal contribution of Tc-svb (Ray et al., in preparation).
Figure Legend Snippet: Schematic representation of Tc-shavenbaby locus ( A ) and transcript ( B ), showing the site at which a GFP-containing marker plasmid was inserted by CRISPR/cas9 genome editing (see also Materials and methods). The mutagenic cassette is inserted into exon 2, that is within the open reading frame upstream of the region encoding the DNA-binding zinc finger domain. Gene disruption leads to mRNA truncation after the insertion site, since Tc-svb expression is absent in homozygous mutants. In addition to segmentation defects and transformation toward thoracic identity, other phenotypes observed in Tc-svb CRISPR mutants include incipient spiracles (possibly a secondary effect of cuticle thinning leading to a defect in the development of tracheal rings); sensory bristles that are shorter and thicker; leg segment boundaries that are not clearly defined; missing leg bristles; unsclerotized pretarsi with soft, rounded apices; and antennae lacking the terminal setae. Therefore, late functions of Tc-svb in epidermal and appendage differentiation are strongly affected in Tc-svb CRISPR mutant embryos, while the segmentation phenotype is milder that Tc-svb -RNAi knockdown due to maternal contribution of Tc-svb (Ray et al., in preparation).

Techniques Used: Marker, Plasmid Preparation, CRISPR, Binding Assay, Expressing, Transformation Assay, Mutagenesis

Related Articles

Clone Assay:

Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning
Article Snippet: .. DNA constructs and transgenics To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP ( ) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech). .. To obtain the pUASp-Svb-3Kmut-GFP, the EcoRI fragment with the 3 K mutated from pAc-SvbK7 ( ) was cloned into the pUASp-Svb::GFP, linearized with EcoRI.

Amplification:

Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning
Article Snippet: .. DNA constructs and transgenics To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP ( ) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech). .. To obtain the pUASp-Svb-3Kmut-GFP, the EcoRI fragment with the 3 K mutated from pAc-SvbK7 ( ) was cloned into the pUASp-Svb::GFP, linearized with EcoRI.

Construct:

Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning
Article Snippet: .. DNA constructs and transgenics To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP ( ) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech). .. To obtain the pUASp-Svb-3Kmut-GFP, the EcoRI fragment with the 3 K mutated from pAc-SvbK7 ( ) was cloned into the pUASp-Svb::GFP, linearized with EcoRI.

Polymerase Chain Reaction:

Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning
Article Snippet: .. DNA constructs and transgenics To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP ( ) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech). .. To obtain the pUASp-Svb-3Kmut-GFP, the EcoRI fragment with the 3 K mutated from pAc-SvbK7 ( ) was cloned into the pUASp-Svb::GFP, linearized with EcoRI.

Transformation Assay:

Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning
Article Snippet: .. DNA constructs and transgenics To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP ( ) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech). .. To obtain the pUASp-Svb-3Kmut-GFP, the EcoRI fragment with the 3 K mutated from pAc-SvbK7 ( ) was cloned into the pUASp-Svb::GFP, linearized with EcoRI.

Plasmid Preparation:

Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning
Article Snippet: .. DNA constructs and transgenics To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP ( ) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech). .. To obtain the pUASp-Svb-3Kmut-GFP, the EcoRI fragment with the 3 K mutated from pAc-SvbK7 ( ) was cloned into the pUASp-Svb::GFP, linearized with EcoRI.

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    TaKaRa puasp svb
    Effect of modified forms of the <t>Svb</t> protein on epidermal trichome formation. <t>UAS-GFP</t> (control) ( A,A’ ), UAS-Svb-ACT ( B,B’ ) and UAS-Svb-3Kmut ( C,C’ ) were expressed in the embryonic epidermis under the control of the ptc-Gal4 driver. Top rows show whole embryo cuticles ( A–C ), the bottom row shows close-ups in the ventral region of the third abdominal segment ( A’–C’ ). Svb-ACT, which lacks the N-terminal repressor domain and thus mimics the processed form of Svb, acts as a constitutive activator of transcription and triggers the production of ectopic trichomes. In contrast, Svb-3Kmut -bearing mutations on the 3 Lysines ubiquitinated by Ubr3 in response to Tal peptides- behaves as a repressor and counteracts endogenous Svb activity, resulting in loss of trichomes.
    Puasp Svb, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puasp svb/product/TaKaRa
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    puasp svb - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    Image Search Results


    Effect of modified forms of the Svb protein on epidermal trichome formation. UAS-GFP (control) ( A,A’ ), UAS-Svb-ACT ( B,B’ ) and UAS-Svb-3Kmut ( C,C’ ) were expressed in the embryonic epidermis under the control of the ptc-Gal4 driver. Top rows show whole embryo cuticles ( A–C ), the bottom row shows close-ups in the ventral region of the third abdominal segment ( A’–C’ ). Svb-ACT, which lacks the N-terminal repressor domain and thus mimics the processed form of Svb, acts as a constitutive activator of transcription and triggers the production of ectopic trichomes. In contrast, Svb-3Kmut -bearing mutations on the 3 Lysines ubiquitinated by Ubr3 in response to Tal peptides- behaves as a repressor and counteracts endogenous Svb activity, resulting in loss of trichomes.

    Journal: eLife

    Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning

    doi: 10.7554/eLife.39748

    Figure Lengend Snippet: Effect of modified forms of the Svb protein on epidermal trichome formation. UAS-GFP (control) ( A,A’ ), UAS-Svb-ACT ( B,B’ ) and UAS-Svb-3Kmut ( C,C’ ) were expressed in the embryonic epidermis under the control of the ptc-Gal4 driver. Top rows show whole embryo cuticles ( A–C ), the bottom row shows close-ups in the ventral region of the third abdominal segment ( A’–C’ ). Svb-ACT, which lacks the N-terminal repressor domain and thus mimics the processed form of Svb, acts as a constitutive activator of transcription and triggers the production of ectopic trichomes. In contrast, Svb-3Kmut -bearing mutations on the 3 Lysines ubiquitinated by Ubr3 in response to Tal peptides- behaves as a repressor and counteracts endogenous Svb activity, resulting in loss of trichomes.

    Article Snippet: DNA constructs and transgenics To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP ( ) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech).

    Techniques: Modification, Activated Clotting Time Assay, Activity Assay

    Schematic representation of Tc-shavenbaby locus ( A ) and transcript ( B ), showing the site at which a GFP-containing marker plasmid was inserted by CRISPR/cas9 genome editing (see also Materials and methods). The mutagenic cassette is inserted into exon 2, that is within the open reading frame upstream of the region encoding the DNA-binding zinc finger domain. Gene disruption leads to mRNA truncation after the insertion site, since Tc-svb expression is absent in homozygous mutants. In addition to segmentation defects and transformation toward thoracic identity, other phenotypes observed in Tc-svb CRISPR mutants include incipient spiracles (possibly a secondary effect of cuticle thinning leading to a defect in the development of tracheal rings); sensory bristles that are shorter and thicker; leg segment boundaries that are not clearly defined; missing leg bristles; unsclerotized pretarsi with soft, rounded apices; and antennae lacking the terminal setae. Therefore, late functions of Tc-svb in epidermal and appendage differentiation are strongly affected in Tc-svb CRISPR mutant embryos, while the segmentation phenotype is milder that Tc-svb -RNAi knockdown due to maternal contribution of Tc-svb (Ray et al., in preparation).

    Journal: eLife

    Article Title: The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning

    doi: 10.7554/eLife.39748

    Figure Lengend Snippet: Schematic representation of Tc-shavenbaby locus ( A ) and transcript ( B ), showing the site at which a GFP-containing marker plasmid was inserted by CRISPR/cas9 genome editing (see also Materials and methods). The mutagenic cassette is inserted into exon 2, that is within the open reading frame upstream of the region encoding the DNA-binding zinc finger domain. Gene disruption leads to mRNA truncation after the insertion site, since Tc-svb expression is absent in homozygous mutants. In addition to segmentation defects and transformation toward thoracic identity, other phenotypes observed in Tc-svb CRISPR mutants include incipient spiracles (possibly a secondary effect of cuticle thinning leading to a defect in the development of tracheal rings); sensory bristles that are shorter and thicker; leg segment boundaries that are not clearly defined; missing leg bristles; unsclerotized pretarsi with soft, rounded apices; and antennae lacking the terminal setae. Therefore, late functions of Tc-svb in epidermal and appendage differentiation are strongly affected in Tc-svb CRISPR mutant embryos, while the segmentation phenotype is milder that Tc-svb -RNAi knockdown due to maternal contribution of Tc-svb (Ray et al., in preparation).

    Article Snippet: DNA constructs and transgenics To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP ( ) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech).

    Techniques: Marker, Plasmid Preparation, CRISPR, Binding Assay, Expressing, Transformation Assay, Mutagenesis