RESUMO
A cDNA encoding a lysozyme was obtained by rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) from females of the malaria vector Anopheles dirus A (Diptera: Culicidae). The 623 bp lysozyme (AdLys c-1) cDNA encodes the 120 amino acid mature protein with a predicted molecular mass of 13.4 kDa and theoretical pI of 8.45. Six cysteine residues and a potential calcium binding motif that are present in AdLys c-1 are highly conserved relative to those of c-type lysozymes found in other insects. RT-PCR analysis of the AdLys c-1 transcript revealed its presence at high levels in the salivary glands both in larval and adult stages and in the larval caecum. dsRNA mediated gene knockdown experiments were conducted to examine the potential role of this lysozyme during Plasmodium berghei infection. Silencing of AdLys c-1 resulted in a significant reduction in the number of oocysts as compared to control dsGFP injected mosquitoes.
Assuntos
Anopheles/genética , Proteínas de Insetos/genética , Insetos Vetores/genética , Malária/transmissão , Muramidase/genética , Glândulas Salivares/metabolismo , Sequência de Aminoácidos , Animais , Anopheles/embriologia , Anopheles/crescimento & desenvolvimento , Sequência Conservada/genética , Cisteína/genética , Feminino , Técnicas de Silenciamento de Genes , Proteínas de Insetos/metabolismo , Insetos Vetores/embriologia , Insetos Vetores/crescimento & desenvolvimento , Larva , Malária/parasitologia , Dados de Sequência Molecular , Muramidase/metabolismo , Plasmodium berghei/fisiologiaRESUMO
A method for rapid determination of viral RNA sequences (RDV) was applied to homogenates of Aedes aegypti collected in Thailand in an area in which dengue fever (dengue hemorrhagic fever) is endemic, using the mosquito cell line C6/36. Nucleic acid sequences of dengue virus type 4 and cell fusing agent virus were detected. This RDV method has the potential to become a standard method for detection of both known and newly emerging, unknown mosquito-borne viruses.
