Dodt G et al. (2000) Biochem J "L-Pipecolic acid oxidase, a human enzyme essential for the degradation of L-pipecolic acid, is most similar to the monomeric sarcosine oxidases."

Reuber BE, Gould SJ, Reimann SA, Kim DG, McCabe K, Dodt G, Mihalik SJ

[Biochem J, 2000]

L-Pipecolic acid oxidase activity is deficient in patients with peroxisome biogenesis disorders (PBDs). Because its role, if any, in these disorders is unknown, we cloned the associated human gene and expressed its protein product. The cDNA was cloned with the use of a reverse genetics approach based on the amino acid sequence obtained from purified L-pipecolic acid oxidase from monkey. The complete cDNA, obtained by conventional library screening and 5' rapid amplification of cDNA ends, encompassed an open reading frame of 1170 bases, translating to a 390-residue protein. The translated protein terminated with the sequence AHL, a peroxisomal targeting signal 1. Indirect immunofluorescence studies showed that the protein product was expressed in human fibroblasts in a punctate pattern that co-localized with the peroxisomal enzyme catalase. A BLAST search with the amino acid sequence showed 31% identity and 53% similarity with Bacillus sp. NS-129 monomeric sarcosine oxidase, as well as similarity to all sarcosine oxidases and dehydrogenases. No similarity was found to the peroxisomal D-amino acid oxidases. The recombinant enzyme oxidized both L-pipecolic acid and sarcosine. However, PBD patients who lack the enzyme activity accumulate only L-pipecolic acid, suggesting that in humans in vivo, this enzyme is involved mainly in the degradation of L-pipecolic acid.

Moss EG (2002) Curr Biol "MicroRNAs: Hidden in the genome."

Moss EG

[Curr Biol, 2002]

Genes for tiny RNAs have been found to be plentiful in the genomes of worms, flies, humans and probably all animals. Some of these microRNAs have been conserved through evolution, and many are expressed only at specific times or places. How they act is just beginning to be understood, but their importance to biology is likely to be great.

Li Y et al. (1997) International C. elegans Meeting "CONTINUING IMPROVEMENTS IN TECHNIQUES FOR PREPARATION OF C. ELEGANS FOR TRANSMISSION ELECTRON MICROSCOPY"

Li Y, Kimble JE

[International C. elegans Meeting, 1997]

We have been continuing to work on protocols for fixation, staining and embedding and want to share our results with the nematode community. Representative micrographs will be shown and protocols will be available for anyone interested.

Dusenbery DB (1985) Comput Biol Med "Using a microcomputer and video camera to simultaneously track 25 animals."

Dusenbery DB

[Comput Biol Med, 1985]

A system that can simultaneously track about 25 animals with the position of each determined once a second is described. The system includes a 6809 microprocessor, OS-9 operating system and application programs written in assembly and BASIC09. The movements and changes in direction of the subjects can be determined and displayed in real time. The system has proven to be valuable in studying the chemotaxis of nematodes and should be applicable to the study of other animals that can be viewed

Yeung MH et al. (1997) International C. elegans Meeting "THE USE OF C. ELEGANS FOR INSECTICIDE DISCOVERY"

Hodgkin JA, Yeung MH, Robinson MP

[International C. elegans Meeting, 1997]

C. elegans can be used as a tool in the discovery of new insecticides. Lead Detection - Wild type C. elegans can be used for screening of unknown compounds. The ideal is to use the target species on its normal host plant. However, that may not be amenable to screening. C. elegans has its advantages: - short life cycle and ease of culturing means the C. elegans can be grown to a large number in a short time using inexpensive culture medium. It can be stored frozen between use. - Our initial study shows that C. elegans has a reasonably selective activity to a spectrum of insecticide standards. Mode Of Action (i) - a panel of C. elegans mutants (each with known resistance or sensitivity) can be used to define the mode of action of unknown compounds. - If a mutant responds in a characteristic way to a compound, it is indicative of a relevant mode of action. - If all mutants do not provide any characteristic response to a compound, it is indicative of a novel mode of action. - Behavioural symptoms can be very useful. - It can be useful for predicting resistance and cross-resistance. - It can be useful for grouping unknown compounds. Mode Of Action (ii) - the unknown compound with a putative novel mode of action can be used as selective agent after EMS mutagenesis to generate mutants. - It can be useful towards understanding the mode of action by carrying out further work on the mutant. - The existing extensive information on ACeDB will facilitate characterisation of the mutant. - The mutant can be screened against a spectrum of putative relevant mode of action insecticide standards to check for cross resistance.

Jansen G et al. (1997) Nat Genet "Reverse genetics by chemical mutagenesis in Caenorhabditis elegans."

Thijssen KL, Plasterk RHA, Hazendonk E, Jansen G

[Nat Genet, 1997]

Traditional reverse genetics on yeast, mice and other organisms uses homologous recombination with transgenic DNA to interrupt a target gene. Here we report that target-selected gene inactivation can be be achieved in Caenorhabditis elegans with the use of chemical mutagens. We use PCR to selectively visualize deletions in genes of interest; the method is sensitive enough to permit detection of a single mutant among more than 15,000 wild types. A permanent frozen mutant collection of more than a million mutagenized animals has been established, and deletion mutants of several G-protein genes were isolated from it. The approach is suitable to be scaled up for systematic inactivation of all 17,000 C. elegans genes. Because it requires no transgenesis or cell culturing, it may also be applicable to small organisms usually considered to be outside the realm of reverse genetics (for example, other nematodes and insects). Any sequenced gene in any organism that can be handled in very large numbers can possibly be targeted in this way.

Wang D et al. (2008) Environ Pollut "Nickel sulfate induces numerous defects in Caenorhabditis elegans that can also be transferred to progeny."

Wang D, Wang Y

[Environ Pollut, 2008]

Whether the multiple biological toxicities from nickel exposure could be transferred to progeny has not been clarified. In this report, we explored the Caenorhabditis elegans to analyze the multiple toxicities of nickel and their possibly transferable properties. The nickel toxicity caused multiple biological defects in a concentration-dependent manner. Moreover, most of these toxicities could be transferred and could be only partially rescued in progeny. Some specific phenotypes in progeny were also found to exhibit no obvious rescue phenotypes or to show even more severe defects than their parents. The defects caused by nickel exposure could be classified into four groups according to their transferring properties. That is, the defects caused by nickel exposure could be largely, or partially, or unable to be rescued, or became even more severe in progeny animals. Therefore, most of the nickel exposure-caused defects can be transferred from parents to their progeny to different degrees in C. elegans.

Fire A et al. (1997) International C. elegans Meeting "SOME PROMOTER ACTIVITY PATTERNS"

Hsieh J, Macarah C, Fire A, Kelly B, Harfe BD, Montgomery MK, Xu SQ

[International C. elegans Meeting, 1997]

We will describe a variety of interesting germline and somatic promoter activity patterns and make some suggestions as to how you might be able to engineer your gene to be expressed in any of all of these patterns.

Buecher EJ et al. (1969) Experientia "Yeast extract as a supplement to chemically defined medium for axenic culture of Caenorhabditis briggsae"

Hansen EL, Buecher EJ

[Experientia, 1969]

Caenorhabditis briggsae, a free living soil nematode, can be grown axenically in a chemically defined medium if this is supplemented with proteinaceous materials. Only a few such materials, derived from chick embryo, bacteria, and liver have been found to be effective supplements. Since yeast cultures on agar support nematodes in vitro it seemed probable that an effective supplement could also be made from extracts of yeast.

Plasterk RHA (2002) Science "RNA silencing: the genome's immune system."

Plasterk RHA

[Science, 2002]

Genomes are databases sensitive to invasion by viruses. In recent years, a defense mechanism has been discovered, which turns out to be conserved among eukaryotes. The system can be compared to the immune system in several ways: It has specificity against foreign elements and the ability to amplify and raise a massive response against an invading nucleic acid. The latter property is beginning to be understood at the molecular level.