Livingstone D (1989) Worm Breeder's Gazette "some more unc-31 sequence."

Livingstone D

[Worm Breeder's Gazette, 1989]

I am continuing the molecular work on the unc-31 gene started by Roger Hoskins. Roger isolated a 17.5kb genomic clone in lambda2001 which he showed was capable of transformation rescue of unc-31 mutants. He also partially sequenced a 4.5kb fragment contained within this clone which covers some of the polymorphisms associated with mutant alleles of the gene. [See Figure 1] I have made subclones of this phage in lambda2001 and have used them in an attempt to narrow down the position of the gene by transformation rescue. A phage containing the 13.5kb XhoI fragment seems to rescue although this result has still to be confirmed. No rescue has been obtained with phage containing the 13.5kb SacI fragment or the 10.5kb SacI-XhoI fragment. I am currently sequencing the 13.5kb rescuing fragment. Several open reading frames have been found which could be spliced together inframe. Some of these have been confirmed by sequencing partial cDNA clones isolated from Julie Ahringers mixed stage cDNA library. The gene covers more than 8kb of genomic DNA. The 5' end is near or beyond the SacI site and the 3' end is beyond the end of the 4.5kb HindIII fragment. No homology has been found between predicted amino acid sequences and protein sequences present in the PIR and Swissprot databases.

Maruyama IN et al. (1986) Worm Breeder's Gazette "a subtle neural connectivity mutant - unc-13."

Nawrocki LW, Maruyama IN

[Worm Breeder's Gazette, 1986]

The loci unc-13 and unc-15 are positioned very close together in the cluster of linkage group I. Both mutants have distinct phenotypes; unc- 15 (e73) worms are limp and paralyzed, while unc-13 (e450) worms are kinked and paralyzed. Mutations at the unc-15 locus affect the putative structural gene for paramyosin. Clones for unc-15 are available and have been used to map a contig that may be long enough to contain the unc-13 locus. A cosmid from the far end of this contig will be used to clone unc-13. Additional incentive to clone unc-13 has come from the isolation of a strain with a restriction site polymorphism from a TR679 background that affects both unc-13 and unc- 15 (see Roger Hoskins' report in this newsletter). In anticipation of cloning unc-13, we wished to identify the lesion causing its phenotype.

Jones D et al. (1992) Worm Breeder's Gazette "A Ubiquitin Fusion Gene from C. elegans"

Candido EPM, Jones D

[Worm Breeder's Gazette, 1992]

Eukaryotes in general have three types of ubiquitin genes: a polyubiquitin gene which encodes multiple tandem copies of ubiquitin transcribed as a polyprotein and two other genes which are fusions of ubiquitin with either a 52 aa or an 80aa ribosomal protein. The polyubiquitin gene from C. elegans was described a few years ago by Roger Graham (Mol. Cell. Biol. (1989)2: 268-271). Since then, we have been studying the regulation of this gene and searching for additional ubiquitin genes. Roger was able to isolate a clone for the 52 aa fusion using a protocol he described in WBG (1990) 11(2):69. A subclone of the ribosomal-protein encoding region was used to probe a YAC grid. A single positive was found, on chromosome III. The most notable aspect of the organization of this gene is the number and position of introns. Two introns occur in the ubiquitin portion of the gene. The position of the first is the same as that in four repeats of the polyubiquitin gene. (C. elegans is the only organism known to have introns in the polyubiquitin gene.) The second intron splits the two terminal glycine residues of ubiquitin. Neither of these intron positions coincides with those in any other 52 aa fusion gene [See Figure 1]. The sequence of a cDNA clone for this gene, isolated from Bob Barstead's lZAP library, shows that the message is trans-spliced. As an additional bonus, this gene has a recognizable TATA motif (unlike the poyubiquitin gene) which is 138 bp from the ATG start codon. We have produced and are currently injecting constructs of this gene. We expect it to be universally constitutive (since it encodes a ribosomal protein) and hope that the promoter may have some practical use. In addition, we may even find out why these ribosomal proteins are always made fused to ubiquitin and what the fate of the ubiquitin part of the fusion is. The search for the third ubiquitin gene continues

Ouazana R (1979) Worm Breeder's Gazette "some enzymatic comparisons between Bergerac and Bristol strains of C elegans."

Ouazana R

[Worm Breeder's Gazette, 1979]

To compare enzymatic activities of Bergerac and Bristol strains of C. elegans, we used the microsystem ' Api Zym ' focused by D. Monget ( 1978) and produced by the french firm API SYSTEM. Some of these tests, not yet commercialized, were a gift of API SYSTEM. The technique consists in microdishes which are impregnated with substrate and buffer ( PH is given in the table ). We introduced 0.1 ml of biological suspension ( a known number of adult worms sonicated in buffer and resuspended in distilled water after homogenization ) in the microdishes which were incubated at 37 C for 4 hours. Bacteria were grossly eliminated by washing; Afterwards their concentration was too low to give response with Api Zym. The reactions were revealed colorimetrically by 4 /oo ' Fast blue BB '. The intensity of the reaction is proportional to enzymatic activity and is noted from - to +++++ . A reaction noted +/- refers to a low but detectable response. To give reliable results, all the tests were read by the same experimenter and enzymatic activities in Bergerac and Bristol strains were performed at the same time. To date, the tests were carried out on 64 enzymatic activities. Most of the enzymes tested were hydrolases ( phosphatases,esterases, aminopeptidases, glycosidases, arylsulfatases ) and some were dehydrogenases. Some of the enzymatic activities tested seem to take a prominent part in postembryonic development. It's the case of Leucine arylamidase, which plays an important part in molting ( Rogers 1965, 1977 ), and could be also the case of glycyl-L-proline arylamidase which is implicated in the metabolism of collagen. The results are summarized in the following table. The two strains show similar enzymatic spectra, but differences can be noted, which are localized essentially on : - Trypsin ( n 8 ) - gamma L-glutamate transpeptidase ( n 38 ) - Glycyl-glycine arylamidase ( n 42 ) - L-threonine arylamidase ( n 54 ) - N-CBZ-glycyl-glycyl-L-arginine arylamidase ( n 56 ) Other differences, in a less degree, but yet detectable, are observed on: - Alkaline Phosphatase ( n 1 ) - L-histidine arylamidase ( n 33 ) - L-leucyl-glycine arylamidase ( n 45 ) - L-ornithine arylamidase ( n 51 ) - L-serine arylamidase ( n 53 ) All the differences observed in this table have been verified in two repetitions, which have similar results. But it could be argued that these differences could be due to nutritional adaptation, because these two strains, xenically cultured on the A1 medium ( for thirty years in case of Bergerac, and one year in the case of Bristol ) are associated with a different bacterial complex. [See Figure 1]

Danielle Thierry-Mieg et al. (2003) Worm Breeder's Gazette "www.wormgenes.org , a new gene centered database"

Vahan Simonyan, Michel Potdevin, Mark Sienkiewicz, Yuji KOHARA, Jean Thierry-Mieg, Danielle Thierry-Mieg, Tadasu SHIN-I

[Worm Breeder's Gazette, 2003]

Wormgenes is a new resource for C.elegans offering a detailed summary about each gene and a powerful query system.

Edgar RS (1975) Worm Breeder's Gazette "the shipping and handling of nematodes."

Edgar RS

[Worm Breeder's Gazette, 1975]

Many of us have totally abandoned wooden sticks for transferring worms. Instead a short (1 - 1 1/2') piece of platinum wire (32 gauge) is sealed in a pasteur pipette holder. Platinum is soft and a sharp hook or spatular end can be shaped with a razor blade and forceps. Various 'tools' of varied shape can be manufactured. Such implements are readily sterilized in a micro-burner. I have had some success shipping worms in letters. Three or four small pieces of filter paper are placed on a piece of aluminum foil. The filter papers are saturated with a buffer washate from a starved plate. The foil is folded several times to create a seal. The recipient places the filter papers on a plate or washes them in buffer.

Gruidl ME et al. (1995) Worm Breeder's Gazette "The Adult Sterile Mutant, let-545, Is Likely To Be a Mutation in glh-1, a Putative P-Granule Component."

Gruidl ME, McKay S, Bennett KL, Rose AM

[Worm Breeder's Gazette, 1995]

The adult sterile mutant, let-545, is likely to be a mutation in glh-1, a putative P-granule component.

Lewis JA (1988) Worm Breeder's Gazette "referencing, crossreferencing, and creating reference lists with Microsoft Word."

Lewis JA

[Worm Breeder's Gazette, 1988]

I've written a little primer on how to use Word (version 3.0 or higher) to automatically generate references (reference number or author-year citation) and cross-references (to page number) within a document using a combination of Word's merge function and table of contents generation facility. Also included in the article are directions for using reference information stored in a database to automatically generate a reference list according to a given style using Word's merge function. The utility of all this is likely to be superseded by future versions of Word and commercially available add- on programs. However, if you are interested, drop me a line and I'll send you a copy of my article.

Way JC et al. (1986) Worm Breeder's Gazette "a simple method for decontaminating worm stocks."

Way JC, Chalfie M

[Worm Breeder's Gazette, 1986]

In the past, we have used the standard method for decontaminating stocks, which involves washing worms off a plate, spinning them in a centrifuge, etc The following procedure is much quicker and requires fewer worms. Also, it can be used to directly decontaminate the progeny of a mating and thus accelerate strain constructions. Using a platinum wire with a glob of bacteria on the bottom, pick up several gravid adults and/or eggs from the contaminated plate and transfer them to a seeded plate. Then put a drop of the usual sodium hypochlorite solution (2-4% NaOCl, .4M NaOH) onto the eggs. About half of the eggs will hatch. Occasionally, a very tough bacterial contaminant will survive to form a few colonies, so it is wise to transfer the survivors on the following day.

Weaver DC et al. (1995) Worm Breeder's Gazette "A Nob Allele at the unc-62 Locus."

Edgar LG, Wood WB, Weaver DC

[Worm Breeder's Gazette, 1995]

A Nob Allele at the unc-62 Locus