Culturing <em>Caenorhabditis elegans</em > in Axenic Liquid Media and Creation of Transgenic Worms by Microparticle Bombardment

Creation of Low-Copy Integrated Transgenic Lines in Caenorhabditis elegans

Genetics ◽

10.1093/genetics/157.3.1217 ◽

2001 ◽

Vol 157 (3) ◽

pp. 1217-1226 ◽

Cited By ~ 1

Author(s):

Vida Praitis ◽

Elizabeth Casey ◽

David Collar ◽

Judith Austin

Keyword(s):

Caenorhabditis Elegans ◽

Expression Level ◽

Dna Arrays ◽

Extrachromosomal Dna ◽

C Elegans ◽

Microparticle Bombardment ◽

Alternative Method ◽

Extrachromosomal Array ◽

Gfp Reporter ◽

Transgenic Lines

Abstract In Caenorhabditis elegans, transgenic lines are typically created by injecting DNA into the hermaphrodite germline to form multicopy extrachromosomal DNA arrays. This technique is a reliable means of expressing transgenes in C. elegans, but its use has limitations. Because extrachromosomal arrays are semistable, only a fraction of the animals in a transgenic extrachromosomal array line are transformed. In addition, because extrachromosomal arrays can contain hundreds of copies of the transforming DNA, transgenes may be overexpressed, misexpressed, or silenced. We have developed an alternative method for C. elegans transformation, using microparticle bombardment, that produces single- and low-copy chromosomal insertions. Using this method, we find that it is possible to create integrated transgenic lines that reproducibly express GFP reporter constructs without the variations in expression level and pattern frequently exhibited by extrachromosomal array lines. In addition, we find that low-copy integrated lines can also be used to express transgenes in the C. elegans germline, where conventional extrachromosomal arrays typically fail to express due to germline silencing.

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A method to rank order water soluble compounds according to their toxicity using Caenorhabditis elegans, a Complex Object Parametric Analyzer and Sorter, and axenic liquid media

Food and Chemical Toxicology ◽

10.1016/j.fct.2009.01.007 ◽

2009 ◽

Vol 47 (4) ◽

pp. 722-728 ◽

Cited By ~ 33

Author(s):

Robert L. Sprando ◽

Nicholas Olejnik ◽

Hediye Nese Cinar ◽

Martine Ferguson

Keyword(s):

Caenorhabditis Elegans ◽

Rank Order ◽

Water Soluble ◽

Complex Object ◽

Liquid Media

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Review for "The connectome of the Caenorhabditis elegans pharynx"

10.1002/cne.24932/v1/review1 ◽

2020 ◽

Author(s):

Sreekanth Chalasani

Keyword(s):

Caenorhabditis Elegans

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The glycomes of Caenorhabditis elegans and other model organisms

Biochemical Society Symposium ◽

10.1042/bss0690117 ◽

2002 ◽

Vol 69 ◽

pp. 117-134 ◽

Cited By ~ 47

Author(s):

Stuart M. Haslam ◽

David Gems ◽

Howard R. Morris ◽

Anne Dell

Keyword(s):

Caenorhabditis Elegans ◽

Current Knowledge ◽

Structural Data ◽

Model Organisms ◽

Entire Genome ◽

C Elegans ◽

The Face ◽

Area Of Concern ◽

Nematode Worm

There is no doubt that the immense amount of information that is being generated by the initial sequencing and secondary interrogation of various genomes will change the face of glycobiological research. However, a major area of concern is that detailed structural knowledge of the ultimate products of genes that are identified as being involved in glycoconjugate biosynthesis is still limited. This is illustrated clearly by the nematode worm Caenorhabditis elegans, which was the first multicellular organism to have its entire genome sequenced. To date, only limited structural data on the glycosylated molecules of this organism have been reported. Our laboratory is addressing this problem by performing detailed MS structural characterization of the N-linked glycans of C. elegans; high-mannose structures dominate, with only minor amounts of complex-type structures. Novel, highly fucosylated truncated structures are also present which are difucosylated on the proximal N-acetylglucosamine of the chitobiose core as well as containing unusual Fucα1–2Gal1–2Man as peripheral structures. The implications of these results in terms of the identification of ligands for genomically predicted lectins and potential glycosyltransferases are discussed in this chapter. Current knowledge on the glycomes of other model organisms such as Dictyostelium discoideum, Saccharomyces cerevisiae and Drosophila melanogaster is also discussed briefly.

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