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    Promega pgemt easy vector
    Localization of <t>Stx2</t> in brains of mice inoculated with pStx2. The area observed in this study is located in the mouse CA2 hippocampus. (A) Brain from control mouse inoculated with <t>pGEMT.</t> (B) Brain from mouse inoculated with 3 µg of pStx2 24 h after inoculation. (C) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation. (D) Brain from mouse injected with 0.5 ng of Stx2 4 days after injection. (E) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation, without primary antibody. (F) Number of Stx2-immunopositive cells in hippocampus per micrograph. Scale bar = 50 µm.
    Pgemt Easy Vector, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 7609 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Localization of Stx2 in brains of mice inoculated with pStx2. The area observed in this study is located in the mouse CA2 hippocampus. (A) Brain from control mouse inoculated with pGEMT. (B) Brain from mouse inoculated with 3 µg of pStx2 24 h after inoculation. (C) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation. (D) Brain from mouse injected with 0.5 ng of Stx2 4 days after injection. (E) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation, without primary antibody. (F) Number of Stx2-immunopositive cells in hippocampus per micrograph. Scale bar = 50 µm.

    Journal: mBio

    Article Title: Promoter Sequence of Shiga Toxin 2 (Stx2) Is Recognized In Vivo, Leading to Production of Biologically Active Stx2

    doi: 10.1128/mBio.00501-13

    Figure Lengend Snippet: Localization of Stx2 in brains of mice inoculated with pStx2. The area observed in this study is located in the mouse CA2 hippocampus. (A) Brain from control mouse inoculated with pGEMT. (B) Brain from mouse inoculated with 3 µg of pStx2 24 h after inoculation. (C) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation. (D) Brain from mouse injected with 0.5 ng of Stx2 4 days after injection. (E) Brain from mouse inoculated with 0.25 µg of pStx2 48 h after inoculation, without primary antibody. (F) Number of Stx2-immunopositive cells in hippocampus per micrograph. Scale bar = 50 µm.

    Article Snippet: The pGEMT Easy vector carrying the sequence of Stx2 was digested and religated in order to delete the active site of Stx2, generating the plasmid pStx2ΔAB ( ) as a Stx2-specific biological activity control.

    Techniques: Mouse Assay, Injection

    (A) Mice displayed a lethal dose-related response to pStx2. Survival rates of mice injected with different doses of pStx2 are shown. Mice injected with pStx2ΔAB or pGEMT were used as negative controls. Mice injected with 1 LD 100 and 3 LD 100 of purified Stx2 were used as positive controls. (B) Stx2 mRNA was detected in the liver. A real-time-PCR curve using cDNA from livers of mice inoculated with pStx2 (dotted line) or pGEMT (gray line) is shown. As a positive control, 50 pg of plasmid pStx2 (bold line) was used. (C) DNA electrophoresis. Lane 1, 50-bp ladder; lane 2. cDNA from livers of mice inoculated with pStx2; lane 3, cDNA from livers of mice inoculated with pGEMT; lane 4, pStx2 was used as positive control.

    Journal: mBio

    Article Title: Promoter Sequence of Shiga Toxin 2 (Stx2) Is Recognized In Vivo, Leading to Production of Biologically Active Stx2

    doi: 10.1128/mBio.00501-13

    Figure Lengend Snippet: (A) Mice displayed a lethal dose-related response to pStx2. Survival rates of mice injected with different doses of pStx2 are shown. Mice injected with pStx2ΔAB or pGEMT were used as negative controls. Mice injected with 1 LD 100 and 3 LD 100 of purified Stx2 were used as positive controls. (B) Stx2 mRNA was detected in the liver. A real-time-PCR curve using cDNA from livers of mice inoculated with pStx2 (dotted line) or pGEMT (gray line) is shown. As a positive control, 50 pg of plasmid pStx2 (bold line) was used. (C) DNA electrophoresis. Lane 1, 50-bp ladder; lane 2. cDNA from livers of mice inoculated with pStx2; lane 3, cDNA from livers of mice inoculated with pGEMT; lane 4, pStx2 was used as positive control.

    Article Snippet: The pGEMT Easy vector carrying the sequence of Stx2 was digested and religated in order to delete the active site of Stx2, generating the plasmid pStx2ΔAB ( ) as a Stx2-specific biological activity control.

    Techniques: Mouse Assay, Injection, Purification, Real-time Polymerase Chain Reaction, Positive Control, Plasmid Preparation, Nucleic Acid Electrophoresis

    The gene las was amplified from Hae11116 strain using the template sequence from GeneBank GI: 14994100. Initially, the upstream region of start codon of las gene was amplified using the primers lasiF and lasiR ( Table 2 ). Also the downstream region of las was amplified with the primers lasfF and lasfR. Both amplicons were cloned in pGEMT Easy (Promega), originating pLAN 75 and pLAN76. In BamHI site of pLAN75, was inserted the ermAM cassette to generate the pLAN77. A new amplification reaction was performed using the ERAM3 and lasiF primers with the pLAN77 as template. The amplicon obtained was inserted in BamHI site of pLAN76 generating the transcriptional fusion vector pLAN78. This vector was then transformed in B4 N. meningitidis strain, originating the LG2 strain.

    Journal: Brazilian Journal of Microbiology

    Article Title: Inflammatory response of Haemophilus influenzae biotype aegyptius causing Brazilian Purpuric Fever

    doi:

    Figure Lengend Snippet: The gene las was amplified from Hae11116 strain using the template sequence from GeneBank GI: 14994100. Initially, the upstream region of start codon of las gene was amplified using the primers lasiF and lasiR ( Table 2 ). Also the downstream region of las was amplified with the primers lasfF and lasfR. Both amplicons were cloned in pGEMT Easy (Promega), originating pLAN 75 and pLAN76. In BamHI site of pLAN75, was inserted the ermAM cassette to generate the pLAN77. A new amplification reaction was performed using the ERAM3 and lasiF primers with the pLAN77 as template. The amplicon obtained was inserted in BamHI site of pLAN76 generating the transcriptional fusion vector pLAN78. This vector was then transformed in B4 N. meningitidis strain, originating the LG2 strain.

    Article Snippet: The lasi and lasf amplicons were cloned in pGEMT Easy (Promega), originating the plasmids pLAN 75 and pLAN76, respectively.

    Techniques: Amplification, Sequencing, Clone Assay, Plasmid Preparation, Transformation Assay

    Construction of transgenic C. elegans expressing Venus under the control of T21D12.3 enhancer/promoter. T21D12.3 and T21D12.4 ( pat-6 ) are transcriptionally regulated as an operon. Therefore, 6.7 kb genome DNA from the transcriptional initiation site to the neighboring gene to pat-6 was subcloned into pGEMT-T plasmid. By injecting the plasmid into germ line of adult worm, we obtained transgenic nematodes in which the Venus cDNA was subcloned into exon 4 to express a PQBP1-Venus fusion protein under the control of endogenous enhancer/promoter.

    Journal: PLoS ONE

    Article Title: Nematode Homologue of PQBP1, a Mental Retardation Causative Gene, Is Involved in Lipid Metabolism

    doi: 10.1371/journal.pone.0004104

    Figure Lengend Snippet: Construction of transgenic C. elegans expressing Venus under the control of T21D12.3 enhancer/promoter. T21D12.3 and T21D12.4 ( pat-6 ) are transcriptionally regulated as an operon. Therefore, 6.7 kb genome DNA from the transcriptional initiation site to the neighboring gene to pat-6 was subcloned into pGEMT-T plasmid. By injecting the plasmid into germ line of adult worm, we obtained transgenic nematodes in which the Venus cDNA was subcloned into exon 4 to express a PQBP1-Venus fusion protein under the control of endogenous enhancer/promoter.

    Article Snippet: The genome fragment was subcloned into pGEMT-T Easy vector (Promega) and the resultant plasmid (pGEMT-T-T21D12.3) was digested with Pml I (BioLabs) which is unique in the exon 4 of the T21D12.3 gene.

    Techniques: Transgenic Assay, Expressing, Plasmid Preparation