WormBase Tree Display for Expr_pattern: Expr2647

Expr2647 Expression_of: Gene: WBGene00004297
    Reflects_endogenous_expression_of: WBGene00004297
  Expression_data: Anatomy_term: WBbt:0005175 Certain
    GO_term: GO:0005634
      GO:0000794
      GO:0043073
  Subcellular_localization: Immunostaining of RAD-51 showed that it forms distinct round or slightly elongated dots within nuclei of the gonad. RAD-51 spots was only occasionally observed in mitotic cells. No RAD-51 staining was observed in rad-51(lg08701) worms or when a pre-immune serum was used. However, RAD-51 foci were readily induced in mitotic cells upon irradiation in a dose- and time-dependent manner. The number of RAD-51 spots was very heterogeneous between cells at comparable stages within one and the same gonad. In a small fraction of nuclei in the mitotic zone 1 or 2 spots occurred. In meiotic cells RAD-51 foci appeared in nuclei of the transition zone of the gonad, which corresponds to the leptotene to zygotene stage. Many of the RAD-51 spots were bar-shaped. In nuclei immediately adjacent to the transition zone (which correspond to late zygotene/early pachytene) up to nine RAD-51 spots were present, whereas by late pachytene they were gone.
  Type: Antibody: Polyclonal rabbit antibody against recombinant protein. A polymerase chain reaction (PCR) fragment encoding the N-terminal 103 amino acids of C. elegans RAD-51 was cloned in frame as a glutathione S-transferase (GST) fusion into pGEX-4T-1 (Amersham) and as a maltose binding protein (MBP) fusion into pMal-c2 (New England BioLabs). Recombinant GST and MPB fusions were expressed in Escherichia coli BL21 CodonPlus (Stratagene), purified under native conditions by affinity chromatography using Glutathione-Sepharose 4B (Amersham) and by amylose-resin affinity chromatography (New England BioLabs). The purified GST protein fusion was dialyzed against phosphate-buffered saline (PBS) and used for immunization of rabbits (Eurogentech). For anti-RAD-51 antibody affinity purification MBP-RAD-51 fusion protein was used. The MBP fusion was dialyzed against 1PBS and 2 mg of total protein was covalently bound to an Affi-Gel 15 matrix (BioRad). Crude serum was diluted 1:1 in 20 mM TRIS, pH 7.5 and pre-cleaned by centrifugation and applied to the column to allow for the binding of the antibody. After washing with 10 mM TRIS-HCl, pH 7.5 and 10 mM TRIS-HCl, pH 7.5, 0.5 mM NaCl, antibodies were eluted with 100 mM glycine, pH 2.5 and the eluates were fractionated and immediately neutralized with 1.5 M TRIS, pH 8.0.
  Pattern: Expressed in gonad.
  Reference: WBPaper00006010
  Antibody_info: WBAntibody00000657