Transfection efficiencies were normalized by using a dual renillaluciferase reporter vector prlsv40. Apr 26, 2016 the pgl3 control vector with the sv40 promoter upstream of the luc gene pgl3 sv40 and the promoterless pgl3 basic vector with the luc gene approx. The reporter vectors are available in four options, depending on your needs. Dependable 80c storage with a focus on sustainability without sacrificing sample protection. The pgl3 bcl2 promotera construct was made by digesting the pgl3 bcl2 promoterfull plasmid with saci. Problem with luciferase reporter assay researchgate.
Isolation and cloning of human nqo1 promoter in pgl3 basic vector asma chinigarzadeh, razauden zulkifli, iman yaze and reyhaneh rahnamai tajadod. You need to be comfortable and find the proper fit, vwr wants to help find the best pipette for you. The reporter vector rv sequencing primers are designed for use with the pgl3 and pgl4 luciferase vectors, chromaluc vectors and pcat3 reporter vectors. Learn more about pgl3 basic vector, 20microg, promega. Basicalimentary tract reg i protein may function as a trophic andor antiapoptotic factor in the development of gastric cancer akira sekikawa, hirokazu fukui, shigehiko fujii,jun takeda. Original article retinoic acid modulates ptgdr promoter activity. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. The originals have been uploaded to and processed by this website, using only the. An expression vector, otherwise known as an expression construct, is usually a plasmid or virus. The pgl3 basic vector lacks eukaryotic promoter and enhancer sequences, allowing maximum flexibility in cloning putative regulatory sequences. In a given assay, 8000 cells from each cell line were plated in designated wells of a 96well plate in triplicate. Jul 15, 2002 to generate the gfp reporter vectors pd2g basic vector, psvd2g promoter vector and psved2g promoterenhancer vector, we replaced the hindiiixbai fragment containing the luciferase gene of pgl3 by the hindiiixbai fragment of pegfp. Paper open access construction and functional analysis of.
The originals have been uploaded to and processed by this website, using only. Promega pgl3 luciferase reporter vectors pgl3 basic vector. Promega corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, highvalue products for the life sciences. Digital collection of empty plasmid backbones from publications and commercially available sources. To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. Basic vectors with no promoter that contain a multiple cloning region for cloning a promoter of choice. Two constructs representing rs383510 genotype tt and genotype cc, as well as unmodified vector pgl3 basic, were transfected into a549 cells. This page is informational only this vector is not available from addgene please contact the manufacturer for further details. Expression vectors are the basic tools in biotechnology for the production of proteins. This listings of restriction sites for the pgl2 luciferase reporter vectors are provided in sections 7.
The saci fragment was subcloned into the pgl3 basic vector digested with saci. Firefly luciferase assays and vectors thermo fisher. The putative enhancer region containing rs383510 was inserted in a luciferase reporter vector. A plant 35s camv promoter induces longterm expression of. Our teams are in regular contact with suppliers and distributors worldwide to manage inventory of raw materials to ensure continued availability. The enhancer vector contains the sv40 enhancer region for putative promoters that may need an additional regulator region. Isolation and cloning of human vegf promoter region in pgl3. The pgl3basic vector lacks eukaryotic promoter and enhancer sequences. Detection of promoter activity by flow cytometric analysis of. Receive the latest news, hot plasmids, discounts and more.
The pgl2basic vector lacks eukaryotic promoter and enhancer sequences. Here are a few examples of the vector magic output for various types of input. This vector is typically used to study upstram or downstream enhancer elements, but not promoters as there is already a promoter, tata box and other elements to run basic transcription. The transfection of the promoter constructs containing the regulatory elements can be performed in many different ways.
Blackwell publishing asiafunctional roles of fli1, a member. Ceph sample na12149a was used as the human genomic template to amplify sequences of interest. It was then inserted into the pgl3 basic vector, which includes luciferase as a reporter gene. Isolation and cloning of human vegf promoter region in pgl3 basic vector iman yaze, razauden zulkifli, asma chinigarzadeh and reyhaneh rahnamai tajadod faculty of bioscience and bioengineering, 810 universiti technologi malaysia abstract worldwide mortality and morbidity from infectious. An example of the use of expression vector is the production of insulin, which is used for. The nfkb1 polymorphism rs4648068 is associated with the.
The backbone of the pgl2 luciferase reporter vectors was redesigned for the pgl3 vectors. Our teams are in regular contact with suppliers and distributors worldwide and are taking all steps necessary to address both demands for diagnostic tools and reliable delivery of all products as quickly as possible. The basic vector has the reporter gene and restriction enzymes sites to clone in a promoter of interest. The pgl3 basic vector was also a kind gift from dr odero. The recombinant plasmids were transformed into li dh5. Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. This is a free resource for the scientific community that is compiled by addgene. We enable science by offering product choice, services, process excellence and our people make it happen. For example, for the empty vector i have luminescence x and for. Nov 03, 2009 the luciferase assay is one of the most wellknown and convenient reporter assays. The pcr products and pgl3basic vector were digested with restriction enzyme xhol thermo and hind.
The recombinant plasmids pgl3 aa and pgl3 gg were constructed for luciferase assay, while the expression plasmids pgl3 aanfkb and pgl3. May 20, 1998 the persistence of transgene expression from pgl3 itr was compared to that of pgl3 by luciferase assays on transiently transfected hela cells as described above and analyzed at 10 days passage 2. Shop online for a wide selection of promega pgl3 luciferase reporter vectors four firefly luciferase reporter vectors. Typically, a promoter region containing 2 to 3 kb of a 5. Firefly luciferase is a 60kda protein produced in nature by several species of the lampyridae family of beetles which includes the genera photinus and luciola. Pcr amplification of the genomic dna is an easy way to obtain the promoter fragment. The originals have been uploaded to and processed by this website, using only the tools made available in the vectorization wizard. If you are using only a small segment of promoter region in pgl3 basic vector, it will not. Realtime quantitative pcr ptgdr, cyp26a1, rara, rarb, rarg, and rxra expression analysis was also performed in basal human. We offer luciferase reporter reagents, multiple cloning site vectors, and flash, glow and dualspectral luciferase assay kits based on the intracellularly expressed firefly luciferase. In particular, promega has developed a firefly luciferase reporter system to study the regulation of gene expression in mammalian cell culture. When transfected into 293t cells, this vector with the cloned tin2 dna fragment was able to drive the expression of the luciferase reporter gene at levels that. Consecutive detection of firefly and renilla luciferase activities in a single sample. Twentyfour hours later, the cells were transiently transfected using.