C. elegans is the first model system that uses green fluorescent protein (GFP) as a visual reporter. Due to its transparency, well-defined anatomy, the multicellular model organism can be examed in vivo non-invasively using GFP. Nowadays, GFP has become one of the standard research tools in C. elegans and is applied extensively in the nematode for observation in vivo of gene expression and cell morphology. Besides, the GFP has also been applied to identify specific subcellular structures via fusing GFP to various proteins. Here we provide the GFP service in C. elegans to suit the particular research needs of our clients.

C. elegans specifically favor the use of GFP

GFP reporter transgenes have become the major tool for expression pattern analysis in C. elegans due to the several characteristics of the worm specifically favor the use of the optical approach. As a simple invertebrate, the nematode is small (<100mm) and transparent, which allows visualization of GFP under a nonconfocal, regular compound microscope as well as a regular dissecting scope equipped with a fluorescent light source. Another advantage of C. elegans is that the construction of transgenic worms expressing a GFP reporter can be easily manipulated and time-saving. On the other hand, there are several advantages of GFP that other reporter gene tags can't replace. GFP has the ability to visualize reporter gene expression in live animals, which allows the visualization of "real time" processes, such as the movement of proteins, organelles, or cells. Besides, this approach provides several days of animal observation if necessary. Moreover, the added advantage is that animals do not have to be sacrificed for visualization of reporter gene expression, for example, the isolation of individual animals with a mutant phenotype in genetic mutant screening.

The application of GFP in C. elegans

• To analyze gene expression patterns
• To determine protein localization and content
• To visualize cellular anatomy
• To assess the development of individual cell types in specific mutant backgrounds- as a cell fate marker
• To measure neuronal function and plasticity
• To identify and isolate cells

Service offerings

• Creation of GFP reporter gene constructs

We offer the in vivo recombination approach to generate GFP reporter gene constructs. This approach relies on the overlapping pieces of DNA injected into C. elegans would undergo efficient homologous recombination (the homologous region is >500 bp). The advantage of this approach is not only the speed, but also the ability to handle virtually unlimited sizes of genomic DNA.

• Germline-mediated transformation by microparticle bombardment

The extrachromosomal arrays created by DNA injected into the gonad of a C. elegans contain multiple copies of the injected DNA. Since the regular arrays are not stably transmitted through mitosis, the high energy irradiation is applied to induce these arrays to become integrated into chromosomes. However, the integration approach can induce mosaicism, such as chromosomally integrated arrays. Here we provide transgenic worms that express GFP fusion proteins through generating "complex arrays". The "complex arrays" are formed through the injection of the desired reporter gene, together with a complex mixture of heterologous DNA. In addition, apart from microinjection, a common technique used for DNA transformation, we also offer microparticle bombardment. The particle bombardment method delivering a low-copy number of the reporter gene benefits the yielding expression levels of the reporter construct, which are more likely to reflect endogenous gene expression levels.

GFP is a commonly used technique and permeates many different research branches in C. elegans. Here we provide GFP service in C. elegans to satisfy customers' particular research needs. CD BioSciences, will be your reliable collaborator. We promise the quality and speed of our service, and also guarantee the satisfaction of our customers. If you have a need in this area of research, please do not hesitate to contact us for more information.

Reference

1. Hobert O, Loria P. et al. (2006). "Uses of GFP in Caenorhabditis elegans". Methods Biochem Anal.;47:203-226.

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