Plasterk RHA (1995) Methods Cell Biol "Reverse genetics: from gene sequence to mutant worm."

Plasterk RHA

[Methods Cell Biol, 1995]

Caenorhabditis elegans is in all likelihood the first metazoan animal whose entire genome will be determined. In addition, a very detailed description of the animal's morphology, development, and physiology is available (see elsewhere in this book, and Wood, 1988). Thus, the complete phenotype and genotype of an animal will be known. What is not known is how genotype determines phenotype; to study this, one needs to establish connections between genome sequence and phenotypes. Much has been done by classic or forward genetics: mutagenesis experiments have identified loci involved in a specific trait. Many of these loci have already been defined at the molecular level, and the genome sequence will certainly aid in the identification of many more. The opposite approach, reverse genetics, becomes naturally more important when more of the genome sequence is determined: Given the sequence of a gene of which nothing else is know, how can the function of that gene be determined? Reverse genetics is more than targeted inactivation. One can study a gene's function by several approaches...|

Fredens J et al. (2014) Methods Mol Biol "Identification of novel protein functions and signaling mechanisms by genetics and quantitative phosphoproteomics in Caenorhabditis elegans."

Larsen MR, Moller-Jensen J, Fredens J, Engholm-Keller K, Faergeman NJ

[Methods Mol Biol, 2014]

Stable isotope labeling by amino acids combined with mass spectrometry is a widely used methodology for measuring relative changes in protein and phosphorylation levels at a global level. We have applied this method to the model organism Caenorhabditis elegans in combination with RNAi-mediated gene knockdown by feeding the nematode on pre-labeled lysine auxotroph Escherichia coli. In this chapter, we describe in details the generation of the E. coli strain, incorporation of heavy isotope-labeled lysine in C. elegans, and the procedure for a comprehensive global phosphoproteomic experiment.

Peredney CL et al. (2000) "Comparison of the toxicological effects of nitrate versus chloride metallic salts on Caenorhabditis elegans in soil."

Williams PL, Peredney CL

[2000]

There is growing interest in the use of bioindicators to assess metal toxicity in soil. The current ASTM Standard Guide for Conducting Laboratory Soil Toxicity Test with the lumbricid earthworm Eisenia fetida (E 1676-97) uses a common earthworm. The nematode Caenorhabditis elegans is a natural soil inhabitant with many characteristics that make an ideal alternate test organism. It has been used to assess metal toxicity in aquatic media, agar plates and in soil. Work is currently underway on the design of a C. elegans procedure for metals in soil. The objective of this study was to determine differences in LC50S between the chloride salt and the nitrate salt forms of cadmium, copper, lead, nickel, and zinc, in three types of soil: Cecil, Tifton, and ASTM artificial soil. Results indicated that the toxicological effect of the metallic salt varies and is dependent on the particular metal. For Cd and Pb the nitrate form is more toxic while Cu and Ni are more toxic in the chloride form. The composition of the soil also effected toxicity, with the metal being the least toxic in ASTM soil and more toxic in the Tifton soil. This strongly correlated with organic matter and clay content of the soil. It is important to determine the effects of carrier salt form and soil composition on metal toxicity, not only in order to standardize the protocol for C. elegans soil toxicity testing, but also in establishing acceptable exposure concentrations in the soil.