Enhancement of green fluorescent protein expression in adeno-associated virus with the woodchuck hepatitis virus post-transcriptional regulatory element.

Abstract

AbstractSince the advent of recombinant DNA technology, maximization of exogenous gene expression has been an important issue for molecular biologists. Efforts at enhancing transgene expression have mostly been directed at improving the efficiency of delivery and increasing levels of transcription and translation. Less progress has been made in the application of post-transcriptional methods for improving gene expression. Here is described the use of an element derived from the woodchuck hepatitis virus (WHV) that possesses the ability to enhance the expression of heterologous genes post-transcriptionally. Green fluorescent protein (GFP) is frequently employed as a fusion protein to enable the detection, visualization, and quantification of molecules under study. Problems of expression are often encountered when using this strategy, making the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) a useful addition to vectors designed to express fusion proteins. This chapter discusses general issues of cloning and placement of the WPRE when designing such vectors.KeywordsGreen Fluorescent ProteinGreen Fluorescent Protein ExpressionEnhance Green Fluorescent Protein ExpressionGreen Fluorescent Protein Fusion ProteinHEPES Buffer SalineThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.