Abstract
Liposome-mediated transfection of cancer cells provide a valuable experimental technique to study cellular gene expression and may also be adapted for gene therapy studies. However, the widely recognized advantage of liposome-mediated transfection is high efficiency. Therefore, this study were performed to optimize transfection techniques in human larynx carcinoma cell line Hep-2 using the commercial synthetic lipid TransFast™ Reagent and monitoring the expression efficiency by using the pSV-?-galactosidase Control Vector which encoded ?-galactosidase, maximum transfection efficiency were achieved with TransFast™ Reagent used at the Charge ratios of 2:1 and 0.5 µg DNA/ml, this is indicate that TransFast™ Reagent can be used as an efficient transfection agent to deliver foreign DNA into human larynx carcinoma cell line Hep-2 and expression of the transgene efficiently.
Highlights
IntroductionThe Escherichia coli lacZ gene methods or reagents in gene transfer encodes β-galactosidase, a tetrameric [9]
The Escherichia coli lacZ gene methods or reagents in gene transfer encodes β-galactosidase, a tetrameric [9].enzyme that catalyzes the hydrolysis ofThe β -galactosidase reporter gene is b-galactoside sugars such as lactose frequently used as a control vector for [1]
A major nucleic acids to eukaryotic cells by a strength of this reporter gene is the nonviral process referred to as ability to assay in situ lipofection, other types of chemical expression with histochemical staining compounds used for transfection
Summary
The Escherichia coli lacZ gene methods or reagents in gene transfer encodes β-galactosidase, a tetrameric [9]. In include: versatility in the addition the widely usage of the gene macromolecules that are delivered, in for studying the activity of promoters vitro and in vivo applications, ability to which is depend on use of cell line for more reproducibly transfect cells that expression [8]. Other studies used this are recalcitrant to other methods, and gene to evaluate the transfection suitability for transient and stable. We use the plasmid vector pSV-ß-galactosidase as an expression vector to evaluate the transfection of Hep-2 cell line using TransFast reagent kit which is a synthetic liposome and monitoring the expression on this cell using β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer kit that depend on spectrophotometric methods and use in situ histochemical analysis using the substrate X-Gal. this will enable us to find the efficiency of TransFast reagent as a DNA delivery vehicle to this type of cell line by determine the optimum values of the Charge ratios of TransFastTM Reagent:DNA and the DNA concentration, we inquire the ability of the these cell for expression foreign DNA which is enable us to transfect the cell with other foreign genes that had therapeutic effect on these kind of cancer cell or to produce any other proteins
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