Abstract
Dot blot hybridization of total RNA from normal and bacteria-injected larvae of the dung beetle Copris tripartitus identified 13 cDNA clones that seem to be associated with the immune response. A cDNA encoding a dung beetle homolog of the heat shock protein HSP70 (CTHSP70) was characterized by full-length sequencing and expression pattern analysis. CTHSP70 was upregulated 4 h after bacterial injection, reached maximum level after 8 h, and declined gradually after 16 h. Our data suggest that CTHSP70 and another 12 genes are involved in the anti-microbial defense that is particularly important for dung beetle larvae living in pathogen-rich conditions.
Highlights
To defend themselves against infection by a variety of bacteria, fungi and parasites, insects have evolved several efficient defense mechanisms, including the production of antimicrobial compounds
The Copris tripartitus larvae were cooled on ice and individually injected with 50 μl of E. coli JM109 (5 × 105 cells) suspended in physiological saline (150 mM NaCl/5 mM KCL)
Construction of the full-length enriched cDNA library cDNA library from larvae of C. tripartitus immunized with E. coli was constructed using a modification of Maruyama and Sugano’s method (Maruyama & Sugano, 1994)
Summary
To defend themselves against infection by a variety of bacteria, fungi and parasites, insects have evolved several efficient defense mechanisms, including the production of antimicrobial compounds. More than 200 antibacterial peptides have been identified in insects. These peptides are known to play important roles in the humoral defense reaction (Hultmark, 2003; Lemaitre, 2004; Hoffmann, 2003; Hoffman et al, 1999; Engstrom, 1999; Garcia-Orozco, 2005). The purpose of the present study was to isolate and identify immune related genes in the dung beetle, C. tripartitus. We cloned immunerelated genes differentially expressed in control and immunized larvae, and detected the time course of expression using Northern blot
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