Chick transfection with GFP

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Green Fluorescent Protein (GFP) as an Indicator of Transfection in Chicken Embryos

Ashley Rowan, F&M College

Introduction

Green fluorescent protein (GFP) is responsible for the bioluminescence of the Pacific Northwest jellyfish, Aequorea victoria. In A.victoria, the 27-kDa protein absorbs blue light from a photoprotein that is activated by calcium and emits green light (2). GFP was isolated from A. victoria and the cDNA was originally cloned in 1992 by Douglas Prasher (1). Genetic manipulations of GFP has made it possible for this protein to be expressed in any organism or cell type(2). GFP expression can be readily detected in a specimen using an ultra violet light source. Because GFP expression can be detected in any organism or cell type, this protein has become a very useful transgenetic tool. Some applications of GFP include: determining transcriptional activity of a gene, cell lineage analyze, development of cell and tissue specific markers, and tracing the pathways and development of pathogens and diseases(2). Embryo transfection is an useful tool for studying molecular signaling as well as cell lineages. For example, transfection techniques can be used to supress signaling by introducing antisense nucleotides into the embryo or ectopic expression can be studied by transfecting the embryo with a retrovirus containing a DNA/cDNA insert of choice. Also, dominant/negative genes can be introduced to study the effect of signal interference on cells.
<http://www.plantsci.cam.ac.uk/Haseloff/GFP/GFPbackgrnd.html>

Figure 1. Aequorea victoria emitting green light.

@Cebra-Thomas, 2001

Last Modified: 2 August 2001

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		Green Fluorescent Protein (GFP) as an Indicator of Transfection in Chicken Embryos
		Ashley Rowan, F&M College
		Introduction
		Green fluorescent protein (GFP) is responsible for the bioluminescence of the Pacific Northwest jellyfish, Aequorea victoria. In A.victoria, the 27-kDa protein absorbs blue light from a photoprotein that is activated by calcium and emits green light (2). GFP was isolated from A. victoria and the cDNA was originally cloned in 1992 by Douglas Prasher (1). Genetic manipulations of GFP has made it possible for this protein to be expressed in any organism or cell type(2). GFP expression can be readily detected in a specimen using an ultra violet light source. Because GFP expression can be detected in any organism or cell type, this protein has become a very useful transgenetic tool. Some applications of GFP include: determining transcriptional activity of a gene, cell lineage analyze, development of cell and tissue specific markers, and tracing the pathways and development of pathogens and diseases(2). Embryo transfection is an useful tool for studying molecular signaling as well as cell lineages. For example, transfection techniques can be used to supress signaling by introducing antisense nucleotides into the embryo or ectopic expression can be studied by transfecting the embryo with a retrovirus containing a DNA/cDNA insert of choice. Also, dominant/negative genes can be introduced to study the effect of signal interference on cells. <http://www.plantsci.cam.ac.uk/Haseloff/GFP/GFPbackgrnd.html> Figure 1. Aequorea victoria emitting green light.