title: Producing a Mammalian GFP Expression Vector Containing Neomycin Resistance Gene.
abstract: The green fluorescent protein (GFP) was originally isolated from the Jellyfish Aequorea Victoria that fluoresces green when exposed to blue light. GFP protein is composed of 238 amino acids with the molecular mass of 26.9 kD. The GFP gene is frequently used in cellular and molecular biology as a reporter gene. To date, many bacterial, yeast, fungal, plants, fly and mammalian cells, including human, have been created which express GFP. Martin Chalfie, Osamu Shimomura, and Roger Tsien were awarded the 2008 noble prize in chemistry for their discovery and development of GFP. In many studies on mammalian cells, GFP gene is introduced into cells using vector-based systems or a recombinant virus to track the location of a target protein or to study the expression level of the gene of interest, but in these studies there is no selection marker to normalize transfection. According to the importance of neomycin gene as a selection marker in mammalian cells, we aimed to produce a GFP expression vector that contains neomycin gene. GFP gene was separated from pEGFP-N1 vector and was inserted in the back-bone of pCDNA3.1/His/LacZ vector that contained the neomycin gene. The resulted vector contained GFP beside neomycin gene.
authors: Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Science, Tehran, Iran.
authors: Manizheh
authors: M
authors: Izadi
authors: Maryam
authors: M
authors: Abiri
authors: Mohammad
authors: M
authors: Keramatipour
keywords: Expression; Green Flourescent Protein; Neomycin; Resistance gene; Vector