| Summary: | 碩士 === 國立陽明大學 === 熱帶醫學研究所 === 93 === In insects, pattern recognition molecules serve as biosenors for detection of invading pathogens in the innate immune system. In this study, a full-length cDNA of Aedes aegypti Gram Negative Binding Protein (AaGNBP) was cloned by Rapid Amplification cDNA End-Polymerase Chain Reaction (RACE-PCR). The AaGNBP cDNA obtained in this study is 1.568 bp with an open reading frame of 503 amino acids bearing a signal peptide and a β1,3-glucanase-like region. The predicted molecular weight of AaGNBP is 57 kDa, but polyclonal antibodies against bacteria-expressed recombinant AaGNBP protein can recognize a 54 kDa peptide in total mosquito lysates, which indicates that a 3 kDa signal peptide may be removed during the processing. Surprisingly, three peptides of 70、72 and 130 kDa, but not 54 kDa, were detected in hemolymph by the same antibodies. It implies that AaGNBPs of hemolymph may be different from what we have. On the other hand, when AaGNBP was silenced by double-strand RNA interference (dsRNAi) in female mosquitoes, it didn’t affect the expression of antibacterial peptides defensin and cecropin. In contrast, it dramatically reduced the melanization efficiency of CM-sephadex in mosquitoes.
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