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. Author manuscript; available in PMC: 2010 Apr 1.
Published in final edited form as: Insect Biochem Mol Biol. 2009 Feb 7;39(4):303–314. doi: 10.1016/j.ibmb.2009.01.007

Figure 2.

Figure 2

Blood-meal-activated expression of transgenic REL2 and Defensin and Cecropin families of AMP in the transgenic Ae. aegypti mosquito strains, REL2-A and REL2-B.

(A) Northern blot analysis: expression profiles of Vitellogenin, endogenous REL2, transgenic REL2, and Defensin A in the fat body of the REL2-A transgenic mosquitoes. Fat bodies were collected from wild-type (WT) and REL2-A mosquitoes: previtellogenic (PV) and blood-fed mosquitoes at 2, 6, 12, 24, and 48 h post blood meal (PBM). WT mosquitoes were injected with E. cloacae, and the fat bodies were collected from non-infected mosquitoes (n) and infected mosquitoes at 2, 6, 12, 24, and 48 h post-infection (PI). Total RNA was extracted and Northern blot analysis performed using the probes for Vg, REL2 RHD domain, and Defensin A; arrows indicate bands for endogenous REL2—REL2 and transgenic REL2—tREL2, (rRNA) ribosomal RNA—loading control.

(B) Northern blot analysis: fat-body-specific expression of transgenic REL2 and Defensin A in REL2-A mosquitoes. Total bodies were collected from previtellogenic (PV) and blood-fed wild-type (WT) and REL2-A mosquitoes at 24 h post blood meal (PBM). Mosquito fat bodies (FB), ovaries (OV), and midguts (MG) were collected from blood-fed mosquitoes at 24 h PBM. Total RNA was extracted, and Northern analysis was performed with the probes for REL2 RHD domain and Defensin A; FB from WT mosquitoes injected with E. cloacae (Ec) at 24 h post-infection were used as a control of Defensin A expression; REL2—endogenous REL2, tREL2—transgenic REL2.

(C) Blood-meal-activated expression of transgenic REL2 and Defensins A, C, and D; and

(D) blood-meal-activated expression of transgenic REL2 and Cecropin A and N in the transgenic REL2-B strain (RT-PCR analysis). Fat bodies were collected from previtellogenic (PV) and blood-fed mosquitoes at 12 and 24 h post blood meal; fat bodies collected from wild-type (WT) mosquitoes injected with E. cloacae (Ec) at 24 h post-infection were used as a control. Total RNA was extracted and treated with DNase I, and cDNA was synthesized and used as a template for RT-PCR with primers specific for transgenic REL2, Defensin A, C, D, and also Cecropin A and N.