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WBPicture0000010178 Description: Figure 5. Cellular expression of the klp-3::lacZ fusion gene in transgenic animals. The scale bar represent 20 um. A, A late 'pretzel' stage embryo showing intense staining in the tail of the late stage embryo. However, no staining is seen in early embryos. B, Nuclear staining of DNA using DAPI dye of the same embryo shown in A. C, Intense staining of the pharyngeal marginal cells in the adult head; anterior is to the left. Notice the absence of staining in the second bulb and the pharyngeal intestinal valve cells. The panel also shows a young L1 larva showing very high staining in the head, and the anterior and posterior region of the larval gut. D, DAPI nuclear staining of the animals shown in C. E, A late larva (L3) showing the intense staining in the pharynx, head and in the posterior gut region. Notice that the staining in the gut has been considerably reduced in later larval stages compared to the early L1 larva, shown in C. F, DAPI staining of the same larval animal shown in E. G, Left dorsolateral view of an adult head. Staining of the possible marginal cells in the pharynx is very intense. H, DAPI staining of the same animal shown in G. Methods: microinjection of the klp-3 genomic fragment and the reporter gene: first a 9 kb KpnI/BamHI fragment, which contains about 3 kb of coding region and 6 kb of upstream promoter, was subcloned from the cosmid T09A5 (obtained from the Sanger Centre) into a pBluescript SK(-) vector. From this 9 kb fragment, a 3 kb HindIII-BglII fragment, which contains 1 kb of upstream promotor and 2 kb of coding region was ligated into the HindIII-BamHI site of the lacZ vector pPD16.51 (Fire et al., 1990). The klp-3::lacZ fusion construct (Figure 2C): for microinjection, the klp-3::lacZ fusion gene was injected with a dominant rol-6 cotransformation marker DNA (kindly given by J. Kramer) at a concentration of 75 ug/ml into the syncitial gonad of the N2 hermaphrodites, as described by Tabish et al. (1995). The him-14 ts(it44) mutant allele was used in the mutant rescue experiments with the klp-3 genomic fragment. Injections were performed at the permissive temperature of 15 C, and transformants were tested at the non-permissive temperature of 25 C. The klp-3::lacZ transformants were histochemically stained and analysed essentially as previously described (Tabish et al., 1995).
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  Depict: Expr_pattern: Expr687
    Anatomy: WBbt:0003673
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  Acknowledgment: Template: WormBase thanks <Journal_URL> for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Reprinted from <Article_URL>. Copyright (<Publication_year>) with permission from <Publisher_URL>.
    Publication_year: 1997
    Article_URL: DOI id 10.1006/jmbi.1997.1112
    Journal_URL: JournalofMolecularBiology
    Publisher_URL: Elsevier
  Reference: WBPaper00002848