Papers by Robert Barstead
Current Opinion in Chemical Biology, Feb 1, 2001
Current Biology, Sep 1, 1999
Journal of Cell Biology, Aug 15, 1991
Humana Press eBooks, Sep 20, 2006
The methods used by the Caenorhabditis elegans Gene Knockout Consortium are conceptually simple. ... more The methods used by the Caenorhabditis elegans Gene Knockout Consortium are conceptually simple. One does a chemical mutagenesis of wild-type C. elegans, and then screens the progeny of the mutagenized animals, in small mixed groups, using polymerase chain reaction (PCR) to identify populations with animals where a portion of DNA bounded by the PCR primers has been deleted. Animals from such populations are then selected and grown clonally to recover a pure genetic strain. We categorize the steps needed to do this as follows: (1) mutagenesis and DNA template preparation, (2) PCR detection of deletions, (3) sibling selection, and (4) deletion stabilization. These are discussed in detail in this chapter.
Molecular Biology of the Cell, Oct 1, 2000
Journal of Molecular Biology, Oct 1, 2003
Briefings in Functional Genomics and Proteomics, Mar 9, 2008
Current Biology, Nov 1, 2000
Proceedings of the National Academy of Sciences of the United States of America, Apr 25, 2000
The EMBO Journal, Oct 1, 1996
Journal of Biological Chemistry, Jun 1, 1989
Molecular Biology of the Cell, Aug 1, 1996
Proceedings of the National Academy of Sciences, 2000
The genome of Caenorhabditis elegans possesses two genes, dpy-18 and phy-2, that encode α subunit... more The genome of Caenorhabditis elegans possesses two genes, dpy-18 and phy-2, that encode α subunits of the enzyme prolyl 4-hydroxylase. We have generated deletions within each gene to eliminate prolyl 4-hydroxylase activity from the animal. The dpy-18 mutant has an aberrant body morphology, consistent with a role of prolyl 4-hydroxylase in formation of the body cuticle. The phy-2 mutant is phenotypically wild type. However, the dpy-18; phy-2 double mutant is not viable, suggesting an essential role for prolyl 4-hydroxylase that is normally accomplished by either dpy-18 or phy-2 . The effects of the double mutation were mimicked by small-molecule inhibitors of prolyl 4-hydroxylase, validating the genetic results and suggesting that C. elegans can serve as a model system for the discovery of new inhibitors.
Molecular Biology of the Cell, 2000
The Caenorhabditis elegans UNC-13 protein and its mammalian homologues are important for normal n... more The Caenorhabditis elegans UNC-13 protein and its mammalian homologues are important for normal neurotransmitter release. We have identified a set of transcripts from the unc-13locus in C. elegans resulting from alternative splicing and apparent alternative promoters. These transcripts encode proteins that are identical in their C-terminal regions but that vary in their N-terminal regions. The most abundant protein form is localized to most or all synapses. We have analyzed the sequence alterations, immunostaining patterns, and behavioral phenotypes of 31 independentunc-13 alleles. Many of these mutations are transcript-specific; their phenotypes suggest that the different UNC-13 forms have different cellular functions. We have also isolated a deletion allele that is predicted to disrupt all UNC-13 protein products; animals homozygous for this null allele are able to complete embryogenesis and hatch, but they die as paralyzed first-stage larvae. Transgenic expression of the entire g...
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Papers by Robert Barstead