Abstract:A transposable element has been isolated from an unstable white mutation in Drosophila mauriliana, a sibling species ofDrosophila melanogaster. The unstable whitepeach (wPch) allele exhibits a spectrum of germ-line and somatic mutability more similar to insertion mutations in maize and in the nematode Caenorhabdits elegans than has been reported for insertion mutations in Drosophila. The We have designated the D. mauritiana transposable element mariner. It was discovered as a repetitive DNA sequence inserted… Show more
“…1 PCR primers used for the amplification of the mariner element from total genomic DNA. The WVPHEL and YSPDLAP regions were determined from the comparison between the mariner-like element (MLE) of Hyalophora cecropia (Lidholm etal., 1991) and the peach element of D. mauritiana (Jacobson et al, 1986). niques was that the washing stringency was generally higher for the former compared with the latter.…”
Section: Resultsmentioning
confidence: 99%
“…For Southern blots, DNA was extracted from about 20-30 flies using the technique of Maruyama & Hartl (1991a). Total DNA was then digested by HindIII and BamHI, two restriction enzymes that do not cut in the peach sequence, an inactive element described in D. mauritiana (Jacobson et al, 1986). Gels were transferred according to the Southern technique (Southern, 1975) and probed with a mixture of pchlV and pch V probes described in Maruyama & Hartl(1991a).…”
Section: Squash Blots and Southern Blotsmentioning
“…1 PCR primers used for the amplification of the mariner element from total genomic DNA. The WVPHEL and YSPDLAP regions were determined from the comparison between the mariner-like element (MLE) of Hyalophora cecropia (Lidholm etal., 1991) and the peach element of D. mauritiana (Jacobson et al, 1986). niques was that the washing stringency was generally higher for the former compared with the latter.…”
Section: Resultsmentioning
confidence: 99%
“…For Southern blots, DNA was extracted from about 20-30 flies using the technique of Maruyama & Hartl (1991a). Total DNA was then digested by HindIII and BamHI, two restriction enzymes that do not cut in the peach sequence, an inactive element described in D. mauritiana (Jacobson et al, 1986). Gels were transferred according to the Southern technique (Southern, 1975) and probed with a mixture of pchlV and pch V probes described in Maruyama & Hartl(1991a).…”
Section: Squash Blots and Southern Blotsmentioning
“…The mariner and Tc1 elements were found in D. mauritiana (Jacobson et al, 1986) and the nematode Caenorhabditis elegans (Emmons et al, 1983), respectively, and related elements are known to occur in diverse taxa, including planarians (Garcia-Fernandez et al, 1995), annelids (Jee et al, 2007), vertebrates (Smit and Riggs, 1996), plants (Jarvik and Lark, 1998) and fungi (Langin et al, 1995). Manifestation of such a wide distribution leads to the inference that elements of this family require no or few factors produced from host cells for their transposition reaction.…”
Tol1 is a DNA-based transposable element residing in the genome of the medaka fish Oryzias latipes, and has been proven to be transposed in various vertebrate species, including mammals. This element belongs to the hAT (hobo/ Activator/Tam3) transposable element family, whose members are distributed in a wide range of organisms. It is thus possible that Tol1 is mobile in organisms other than vertebrates. We here show that transposition of this element occurs in the nematode Caenorhabditis elegans. A donor plasmid containing a Tol1 element and a helper plasmid carrying the transposase gene were delivered into gonad cells and, after several generations of culturing, were recovered from worms. PCR analysis of the donor plasmid, using primers that encompassed the Tol1 element, revealed excision of the Tol1 portion from the plasmid. Analysis of genomic DNA of the worms by the inverse PCR method provided evidence that Tol1 had been integrated into the C. elegans chromosomes. Vertebrates and C. elegans are phylogenetically distantly related organisms in that the former are deuterostomes and the latter a protostome animal. Our results indicate (1) the transposition reaction of the Tol1 element requires, besides the transposase, no factors from host cells, or (2) the host factors, even if required, are those that are common to protostomes and deuterostomes. The results also have significance for the development of a gene transfer vector and other biotechnology tools for C. elegans.
“…15) in the germ line of C. elegans 16,17 .This technique has been used successfully in forward genetic screens to identify genes of interest [18][19][20][21] . This system has three main advantages.…”
We describe a protocol for mutating genes in the nematode Caenorhabditis elegans using the Mos1 transposon of Drosophila mauritiana. Mutated genes containing a Mos1 insertion are molecularly tagged by this heterologous transposable element. Mos1 insertions can therefore be identified in as little as 3 weeks using only basic molecular biology techniques. Mutagenic efficiency of Mos1 is tenfold lower than classical chemical mutagens. However, the ease and speed with which mutagenic insertions can be mapped compares favorably with the vast amount of work involved in classical genetic mapping. Therefore, Mos1 could be the tool of choice when screening procedures are efficient. In addition, Mos1 mutagenesis can greatly simplify the mapping of mutations that exhibit low penetrance, subtle or synthetic phenotypes. The recent development of targeted engineering of C. elegans loci carrying Mos1 insertions further increases the attractiveness of Mos1-mediated mutagenesis.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.