Journal: Insect biochemistry and molecular biology

Article Title: Characterization of a Novel Manduca sexta beta-1, 3-glucan recognition protein (βGRP3) with Multiple Functions

doi: 10.1016/j.ibmb.2014.06.003

Figure Lengend Snippet: (A) Tissue distribution of βGRP3 mRNA. Expression of βGRP3 mRNA in hemocytes, fat body, midgut, epidermis and testis of the 5th instar M. sexta naïve larvae was determined by real-time PCR and normalized to rpS3 gene. (B–D): Expression of βGRP3 mRNA in fat body (B), hemocytes (C), and midgut (D) after immune challenge. Expression of βGRP3 mRNA in hemocytes, fat body and midgut of the 5th instar M. sexta naïve larvae or larvae injected with H2O, E. coli, S. marcescens, S. aureus, B. subtilis, or S. cerevisiae at 24 h post-injection was determined by real-time PCR. The bars represent the mean of three individual measurements ± SEM. Relative expression of βGRP3 mRNA in naïve larval hemocytes (A), or in tissues of naïve larvae (B–D) was set as 1. Comparing expression of βGRP3 mRNA in different tissues (A) or after microbial injections, identical letters among tissues or treatments indicate not significant difference (p>0.05), while different letters indicate significant difference (p<0.05) determined by one way ANOVA followed by a Tukey’s multiple comparison test.

Article Snippet: Significance of difference was determined by one way ANOVA followed by a Tukey’s multiple comparison test using the same software (GraphPad, CA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Injection, Comparison

Journal: Insect biochemistry and molecular biology

Article Title: Characterization of a Novel Manduca sexta beta-1, 3-glucan recognition protein (βGRP3) with Multiple Functions

doi: 10.1016/j.ibmb.2014.06.003

Figure Lengend Snippet: Antibacterial activity of recombinant βGRP3 was determined by inhibition zone assay (A–D) and broth micro-dilution assay (E) as described in the Materials and Methods. Buffer and different amounts of ampicillin (Amp) were used as controls in the inhibition zone assay, the plates were incubated at 37°C overnight, and the diameters of inhibition zones were measured and recorded (A–D). Diluted B. cereus culture was incubated with recombinant βGRP3 alone (2 μg/ml) or βGRP3 (2 μg/ml) mixed with different free microbial components (20 μg/ml each) in 96-well plates at 37°C, and OD600 was recorded every hour up to 22 h. The points represent the mean of four individual measurements ± SEM. Asterisks indicate significant difference in OD600 values at 20 h among the control, βGRP3, and βGRP3+PG-K12 groups (p<0.001) determined by one way ANOVA followed by a Tukey’s multiple comparison test.

Article Snippet: Significance of difference was determined by one way ANOVA followed by a Tukey’s multiple comparison test using the same software (GraphPad, CA).

Techniques: Activity Assay, Recombinant, Inhibition, Microdilution Assay, Incubation, Control, Comparison