Isolation and Characterization of a Defensin-Like Peptide (Coprisin) from the Dung Beetle, Copris tripartitus
Figure 1
Isolation of Coprisin cDNA. (a) Differential dot blot hybridization. Randomly selected 1862 cDNA clones from cDNA library of Copris tripartitus larvae were hybridized with probes of E. coli-immunized larvae. In this figure, one pair of hybridization dot blots is shown for the Coprisin cDNA clone (adapted from Hwang et al. [13]). The arrow indicates a differentially expressed Coprisin cDNA clone in saline- or E. coli-immunized larvae. (b) Northern blot hybridization of the Coprisin gene for a time-course after immunization. C. tripartitus larvae were injected with 50 L of E. coli JM109 ( cells) suspended in physiological saline (150 mM NaCl/5 mM KCl). Larvae were kept for 0, 4, 8, 16, and 24 hours. Ten microgram aliquots of total RNA were resolved on formaldehyde containing agarose gels and blotted onto nitrocellulose membranes. The probe was labeled with [-32P]dCTP. As an internal marker, 28S rRNA was stained with ethidium bromide.
