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Promega plasmid p gl3-basic
Plasmid P Gl3 Basic, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/plasmid+p+gl3-basic/gl3+basic+vector/pmc11739467-458-21-24
Average 90 stars, based on 1 article reviews
plasmid p gl3-basic - by Bioz Stars, 2026-09
90/100 stars

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Article Title: Oxindole-curcumin hybrid compound enhances the transcription of γ-glutamylcysteine ligase.
Article Snippet: The PCR-amplified products were subcloned into GL3-Basic (Promega Corporation, Madison, WI, USA) at MluI and XhoI restriction sites.

Article Title: Clinical and virological implications of A1846T and C1913A/G mutations of hepatitis B virus genome in severe liver diseases.
Article Snippet: Objective: Mutations occurring within different genes of hepatitis B virus (HBV) genome may have different clinical implications.. This study aimed to observe the clinical and virological implications of the A1846T and C1913A/G mutations of HBV genome in the development and treatment outcome of severe liver diseases, which has not been previously determined.. Materials and methods: A total of 438 cases of patients with liver diseases were retrospectively reviewed, including 146 with mild chronic hepatitis B infection (CHB-M), 146 with severe chronic hepatitis B infection (CHB-S), and 146 with acute-on-chronic liver failure (ACLF).

Article Title: An atlas of neuropathic pain-associated molecular pathological characteristics in the mouse spinal cord
Article Snippet: To identify the promoter activity of Penk , HEK293T cells cultured in the 12-well culture plates were transfected with 0.5 μg plasmid p GL3-basic (Promega, Madison, WI, USA) or p GL3- Penk -promoter constructed by Genecreate (Wuhan Genecreate Biotechnology Engineering Co., Ltd., Wuhan, China) that encompasses a 1500 bp region preceding the transcription start site.

Article Title: Thyroid-stimulating hormone improves insulin sensitivity in skeletal muscle cells via cAMP/PKA/CREB pathway-dependent upregulation of insulin receptor substrate-1 expression.
Article Snippet: Thyroid-stimulating hormone (TSH) receptor is expressed in extrathyroidal tissues such as hepatocytes, adipocytes, and skeletal muscle, which suggests a possible novel role of TSH in various metabolic processes in extrathyroidal tissues independent of thyroid hormones.. We investigated whether TSH has any effects on glucose tolerance and insulin sensitivity in the skeletal muscle using diet-induced obesity (DIO) mouse models and rodent skeletal muscle cells.. TSH improved glucose tolerance in DIO mice and this was associated with an improvement of skeletal muscle insulin sensitivity resulting from the increased expression of insulin receptor substrate (IRS)-1 protein and mRNA therein.

Luciferase:

Article Title: E2F4-induced AGAP2-AS1 up-regulation accelerates the progression of colorectal cancer via miR-182-5p/CFL1 axis.
Article Snippet: Background: Long non-coding RNAs (lncRNAs) are closely associated with the pathogenesis of numerous diseases including cancers.. LncRNA AGAP2 Antisense RNA 1 (AGAP2-AS1) has been found to participate in the tumorigenesis of several kinds of human cancers.. Nonetheless, its potential function in colorectal cancer (CRC) was still poorly investigated.

Reporter Assay:

Article Title: E2F4-induced AGAP2-AS1 up-regulation accelerates the progression of colorectal cancer via miR-182-5p/CFL1 axis.
Article Snippet: Background: Long non-coding RNAs (lncRNAs) are closely associated with the pathogenesis of numerous diseases including cancers.. LncRNA AGAP2 Antisense RNA 1 (AGAP2-AS1) has been found to participate in the tumorigenesis of several kinds of human cancers.. Nonetheless, its potential function in colorectal cancer (CRC) was still poorly investigated.

Synthesized:

Article Title: E2F4-induced AGAP2-AS1 up-regulation accelerates the progression of colorectal cancer via miR-182-5p/CFL1 axis.
Article Snippet: Background: Long non-coding RNAs (lncRNAs) are closely associated with the pathogenesis of numerous diseases including cancers.. LncRNA AGAP2 Antisense RNA 1 (AGAP2-AS1) has been found to participate in the tumorigenesis of several kinds of human cancers.. Nonetheless, its potential function in colorectal cancer (CRC) was still poorly investigated.

Clone Assay:

Article Title: E2F4-induced AGAP2-AS1 up-regulation accelerates the progression of colorectal cancer via miR-182-5p/CFL1 axis.
Article Snippet: Background: Long non-coding RNAs (lncRNAs) are closely associated with the pathogenesis of numerous diseases including cancers.. LncRNA AGAP2 Antisense RNA 1 (AGAP2-AS1) has been found to participate in the tumorigenesis of several kinds of human cancers.. Nonetheless, its potential function in colorectal cancer (CRC) was still poorly investigated.

Article Title: Long non-coding RNA H19 and the underlying epigenetic function in response to DNA damage of lung cancer cells
Article Snippet: .. Next, the WT or MUT of H19-binding miR-675 (or miR-625-5p) was cloned intopGL3 Basic vector (Promega, Madison, WI, USA). ..

Article Title: Identification and functional characterization of a glucose regulated protein 94 gene in Litopenaeus vannamei and its responsiveness in WSSV infection.
Article Snippet: In the current study, a cDNA of glucose regulated protein 94 (LvGRP94) was cloned from Litopenaeus vannamei.. Subcellular localization assay revealed that LvGRP94 expressed in endoplasmic reticulum (ER).. And results of reported gene assays demonstrated that the promoter of LvGRP94 was activated by L. vannamei leucine zipper domain transcription factor X-box binding protein 1 (LvXBP1) or heat shock treatment.

Article Title: Expression of transcription factors KLF2 and KLF4 is induced by the mammalian Golgi stress response
Article Snippet: The plasmids expressing human KLF family proteins were purchased from Genscript (Piscataway, NJ) or OriGene (Rockville, MD). .. To construct promoter-luciferase reporter vectors, the corresponding promoter regions of the human KLF2 and KLF4 genes were amplified using human genomic DNA as a template and cloned into the BglII site of the GL4 basic vector (Promega, Fitchburg, WI). .. Point mutants of the vectors were constructed by site-directed mutagenesis using a QuikChange Site-Directed Mutagenesis kit (Stratagene, CA) ( Yoshida et al ., 2006 ).

Plasmid Preparation:

Article Title: Long non-coding RNA H19 and the underlying epigenetic function in response to DNA damage of lung cancer cells
Article Snippet: .. Next, the WT or MUT of H19-binding miR-675 (or miR-625-5p) was cloned intopGL3 Basic vector (Promega, Madison, WI, USA). ..

Article Title: Identification and functional characterization of a glucose regulated protein 94 gene in Litopenaeus vannamei and its responsiveness in WSSV infection.
Article Snippet: In the current study, a cDNA of glucose regulated protein 94 (LvGRP94) was cloned from Litopenaeus vannamei.. Subcellular localization assay revealed that LvGRP94 expressed in endoplasmic reticulum (ER).. And results of reported gene assays demonstrated that the promoter of LvGRP94 was activated by L. vannamei leucine zipper domain transcription factor X-box binding protein 1 (LvXBP1) or heat shock treatment.

Article Title: Expression of transcription factors KLF2 and KLF4 is induced by the mammalian Golgi stress response
Article Snippet: The plasmids expressing human KLF family proteins were purchased from Genscript (Piscataway, NJ) or OriGene (Rockville, MD). .. To construct promoter-luciferase reporter vectors, the corresponding promoter regions of the human KLF2 and KLF4 genes were amplified using human genomic DNA as a template and cloned into the BglII site of the GL4 basic vector (Promega, Fitchburg, WI). .. Point mutants of the vectors were constructed by site-directed mutagenesis using a QuikChange Site-Directed Mutagenesis kit (Stratagene, CA) ( Yoshida et al ., 2006 ).

Polymerase Chain Reaction:

Article Title: Identification and functional characterization of a glucose regulated protein 94 gene in Litopenaeus vannamei and its responsiveness in WSSV infection.
Article Snippet: In the current study, a cDNA of glucose regulated protein 94 (LvGRP94) was cloned from Litopenaeus vannamei.. Subcellular localization assay revealed that LvGRP94 expressed in endoplasmic reticulum (ER).. And results of reported gene assays demonstrated that the promoter of LvGRP94 was activated by L. vannamei leucine zipper domain transcription factor X-box binding protein 1 (LvXBP1) or heat shock treatment.

Amplification:

Article Title: Identification and functional characterization of a glucose regulated protein 94 gene in Litopenaeus vannamei and its responsiveness in WSSV infection.
Article Snippet: In the current study, a cDNA of glucose regulated protein 94 (LvGRP94) was cloned from Litopenaeus vannamei.. Subcellular localization assay revealed that LvGRP94 expressed in endoplasmic reticulum (ER).. And results of reported gene assays demonstrated that the promoter of LvGRP94 was activated by L. vannamei leucine zipper domain transcription factor X-box binding protein 1 (LvXBP1) or heat shock treatment.

Article Title: Expression of transcription factors KLF2 and KLF4 is induced by the mammalian Golgi stress response
Article Snippet: The plasmids expressing human KLF family proteins were purchased from Genscript (Piscataway, NJ) or OriGene (Rockville, MD). .. To construct promoter-luciferase reporter vectors, the corresponding promoter regions of the human KLF2 and KLF4 genes were amplified using human genomic DNA as a template and cloned into the BglII site of the GL4 basic vector (Promega, Fitchburg, WI). .. Point mutants of the vectors were constructed by site-directed mutagenesis using a QuikChange Site-Directed Mutagenesis kit (Stratagene, CA) ( Yoshida et al ., 2006 ).

Construct:

Article Title: Identification and functional characterization of a glucose regulated protein 94 gene in Litopenaeus vannamei and its responsiveness in WSSV infection.
Article Snippet: In the current study, a cDNA of glucose regulated protein 94 (LvGRP94) was cloned from Litopenaeus vannamei.. Subcellular localization assay revealed that LvGRP94 expressed in endoplasmic reticulum (ER).. And results of reported gene assays demonstrated that the promoter of LvGRP94 was activated by L. vannamei leucine zipper domain transcription factor X-box binding protein 1 (LvXBP1) or heat shock treatment.

Article Title: Expression of transcription factors KLF2 and KLF4 is induced by the mammalian Golgi stress response
Article Snippet: The plasmids expressing human KLF family proteins were purchased from Genscript (Piscataway, NJ) or OriGene (Rockville, MD). .. To construct promoter-luciferase reporter vectors, the corresponding promoter regions of the human KLF2 and KLF4 genes were amplified using human genomic DNA as a template and cloned into the BglII site of the GL4 basic vector (Promega, Fitchburg, WI). .. Point mutants of the vectors were constructed by site-directed mutagenesis using a QuikChange Site-Directed Mutagenesis kit (Stratagene, CA) ( Yoshida et al ., 2006 ).



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HNF4α regulates the promoter of the RARβ gene in HepG2 cells. HepG2 cells in 24-well plates were co-transfected for 24 h with a fragment of the human RARβ promoter expanding from −1.7 kbp from transcription start site to +0.217 kpb, as the full length promoter construct, in p <t>GL3-basic-luc</t> vector together with p RLTK plasmid containing Renilla-luc, as the control, and with either human HNF4α, RARα/RXRα, or all three transcription factors, and then treated further either without ( A ) or with RA for 1 to 24 h, after which the cells were collected to assay for luciferase activity. HNF4α alone or with RARα/RXRα suppressed the promoter activity of the human RARβ gene in the cells treated without RA ( A ), but upregulated the promoter when the cells were treated with RA following transfection ( B ). HNF4α regulates the promoter of the mouse RARβ gene in HepG2 cells ( C ). It suppresses the promoter activity of the mouse gene (empty bars in C) in HepG2 cells treated with the vehicle, but it increases the promoter activity in the cells treated with RA (black bar in C).
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HNF4α regulates the promoter of the RARβ gene in HepG2 cells. HepG2 cells in 24-well plates were co-transfected for 24 h with a fragment of the human RARβ promoter expanding from −1.7 kbp from transcription start site to +0.217 kpb, as the full length promoter construct, in p GL3-basic-luc vector together with p RLTK plasmid containing Renilla-luc, as the control, and with either human HNF4α, RARα/RXRα, or all three transcription factors, and then treated further either without ( A ) or with RA for 1 to 24 h, after which the cells were collected to assay for luciferase activity. HNF4α alone or with RARα/RXRα suppressed the promoter activity of the human RARβ gene in the cells treated without RA ( A ), but upregulated the promoter when the cells were treated with RA following transfection ( B ). HNF4α regulates the promoter of the mouse RARβ gene in HepG2 cells ( C ). It suppresses the promoter activity of the mouse gene (empty bars in C) in HepG2 cells treated with the vehicle, but it increases the promoter activity in the cells treated with RA (black bar in C).

Journal: International Journal of Molecular Sciences

Article Title: Hepatocyte Nuclear Factor 4α (HNF4α) Plays a Controlling Role in Expression of the Retinoic Acid Receptor β ( RARβ ) Gene in Hepatocytes

doi: 10.3390/ijms24108608

Figure Lengend Snippet: HNF4α regulates the promoter of the RARβ gene in HepG2 cells. HepG2 cells in 24-well plates were co-transfected for 24 h with a fragment of the human RARβ promoter expanding from −1.7 kbp from transcription start site to +0.217 kpb, as the full length promoter construct, in p GL3-basic-luc vector together with p RLTK plasmid containing Renilla-luc, as the control, and with either human HNF4α, RARα/RXRα, or all three transcription factors, and then treated further either without ( A ) or with RA for 1 to 24 h, after which the cells were collected to assay for luciferase activity. HNF4α alone or with RARα/RXRα suppressed the promoter activity of the human RARβ gene in the cells treated without RA ( A ), but upregulated the promoter when the cells were treated with RA following transfection ( B ). HNF4α regulates the promoter of the mouse RARβ gene in HepG2 cells ( C ). It suppresses the promoter activity of the mouse gene (empty bars in C) in HepG2 cells treated with the vehicle, but it increases the promoter activity in the cells treated with RA (black bar in C).

Article Snippet: Construction of the plasmid vectors including p GL3-Basic-hCYP26A1-E4-luc (submitted to addgene.org), p GL3-Basic-hRARβp-luc (human RARβ2 promoter), p GL3-Basic-hCYP2C9p-luc, p cDNNA3.1-hRARα.hRXRα (submitted to addgene.org), and p cDNA3.1-hHNF4α were reported previously [ , , , , ].

Techniques: Transfection, Construct, Plasmid Preparation, Control, Luciferase, Activity Assay

Mutation of the critical residues present in the ligand binding domain of human HNF4α suppresses transcription activation of the promoters of human APOC3 and CYP2C9 genes. HepG2 cells were co-transfected with either the p GL3-b-hApoC3 ( A – C ) or p GL3-b-hCYP2C9 ( D – F ) promoter construct together with either hHNF4α, RARα/RXRα, or all three transcription factors, and then treated with either vehicle or 1 µM RA for 24 h, after which the cells were assayed for their luciferase activity. The effects of the individual mutant residues in the ligand binding domain of the human HNF4α compared with WT HNF4α on the promoter activity of APOC3 ( B ) or CYP2C9 ( E ) in HepG2 cells treated with either vehicle or 1 μM RA for 24 h. The endogenous HNF4α transcriptional activity toward APOC3 ( C ) and CYP2C9 ( F ) promoters was assessed in HepG2 cells with or without the addition of human HNF4α mutants. Data from each bar represent the mean of n = 3 wells ± SD. HNF4α mutant #’s are as follows: (1) S190K, (2) 191MK, (3) R221G, (4) L228K, (5) L229K, (6) R235G, (7) I347K, and (8) I355K.

Journal: International Journal of Molecular Sciences

Article Title: Hepatocyte Nuclear Factor 4α (HNF4α) Plays a Controlling Role in Expression of the Retinoic Acid Receptor β ( RARβ ) Gene in Hepatocytes

doi: 10.3390/ijms24108608

Figure Lengend Snippet: Mutation of the critical residues present in the ligand binding domain of human HNF4α suppresses transcription activation of the promoters of human APOC3 and CYP2C9 genes. HepG2 cells were co-transfected with either the p GL3-b-hApoC3 ( A – C ) or p GL3-b-hCYP2C9 ( D – F ) promoter construct together with either hHNF4α, RARα/RXRα, or all three transcription factors, and then treated with either vehicle or 1 µM RA for 24 h, after which the cells were assayed for their luciferase activity. The effects of the individual mutant residues in the ligand binding domain of the human HNF4α compared with WT HNF4α on the promoter activity of APOC3 ( B ) or CYP2C9 ( E ) in HepG2 cells treated with either vehicle or 1 μM RA for 24 h. The endogenous HNF4α transcriptional activity toward APOC3 ( C ) and CYP2C9 ( F ) promoters was assessed in HepG2 cells with or without the addition of human HNF4α mutants. Data from each bar represent the mean of n = 3 wells ± SD. HNF4α mutant #’s are as follows: (1) S190K, (2) 191MK, (3) R221G, (4) L228K, (5) L229K, (6) R235G, (7) I347K, and (8) I355K.

Article Snippet: Construction of the plasmid vectors including p GL3-Basic-hCYP26A1-E4-luc (submitted to addgene.org), p GL3-Basic-hRARβp-luc (human RARβ2 promoter), p GL3-Basic-hCYP2C9p-luc, p cDNNA3.1-hRARα.hRXRα (submitted to addgene.org), and p cDNA3.1-hHNF4α were reported previously [ , , , , ].

Techniques: Mutagenesis, Ligand Binding Assay, Activation Assay, Transfection, Construct, Luciferase, Activity Assay

Mutation of the critical amino acid residues present in the ligand binding domain of human HNF4α suppresses transcription activation of the promoters of human CYP26A1 and RARβ genes in HepG2 cells treated with RA. HepG2 cells grown in 24-well plates were co-transfected with either p GL3-b-hCYP26A1 ( A ) or p GL3-b-hRARβ ( B ) promoters, each with p RLTK as the control, together with either wildtype (WT) HNF4α or its individual mutants (Mutant # 1 to 8), and then treated with either vehicle or 1 µM RA for 24 h, after which the cells were assayed for luciferase activity. Data from each bar represent the mean of n = 3 wells ± SD. HNF4α mutant #’s are as follows: (1) S190K, (2) 191MK, (3) R221G, (4) L228K, (5) L229K, (6) R235G, (7) I347K, and (8) I355K.

Journal: International Journal of Molecular Sciences

Article Title: Hepatocyte Nuclear Factor 4α (HNF4α) Plays a Controlling Role in Expression of the Retinoic Acid Receptor β ( RARβ ) Gene in Hepatocytes

doi: 10.3390/ijms24108608

Figure Lengend Snippet: Mutation of the critical amino acid residues present in the ligand binding domain of human HNF4α suppresses transcription activation of the promoters of human CYP26A1 and RARβ genes in HepG2 cells treated with RA. HepG2 cells grown in 24-well plates were co-transfected with either p GL3-b-hCYP26A1 ( A ) or p GL3-b-hRARβ ( B ) promoters, each with p RLTK as the control, together with either wildtype (WT) HNF4α or its individual mutants (Mutant # 1 to 8), and then treated with either vehicle or 1 µM RA for 24 h, after which the cells were assayed for luciferase activity. Data from each bar represent the mean of n = 3 wells ± SD. HNF4α mutant #’s are as follows: (1) S190K, (2) 191MK, (3) R221G, (4) L228K, (5) L229K, (6) R235G, (7) I347K, and (8) I355K.

Article Snippet: Construction of the plasmid vectors including p GL3-Basic-hCYP26A1-E4-luc (submitted to addgene.org), p GL3-Basic-hRARβp-luc (human RARβ2 promoter), p GL3-Basic-hCYP2C9p-luc, p cDNNA3.1-hRARα.hRXRα (submitted to addgene.org), and p cDNA3.1-hHNF4α were reported previously [ , , , , ].

Techniques: Mutagenesis, Ligand Binding Assay, Activation Assay, Transfection, Control, Luciferase, Activity Assay