RESUMO
Prenyltransferases play a key role in isoprenoid biosynthesis. Here, a cDNA encoding a prenyltransferase was isolated from the cotton aphid, Aphis gossypii, which consists of 1354 nucleotides and encodes a protein of 394 amino acids (AgIPPS). Subsequent sequencing of AgIPPS genomic DNA resulted in one 3138-bp sequence. Southern blotting analysis indicated that only a single IPPS gene exists in the cotton aphid. Real-time quantitative PCR showed that AgIPPS transcripts were mainly present at the corpora allatum, but small quantity could be detected in tissues other than the corpora allatum. Transcript abundance changed in an alternative manner at different life stages. High expression was observed in embryos, second and forth instar nymphs and adults, but only low level was detected in the first and third instars. Functional expression, activity assay and product analysis revealed that the mature form of AgIPPS (AgIPPS-S) could efficiently convert labeled isopentenyl diphosphate in the presence of dimethylallyl diphosphate to both geranyl diphosphate (GPP) and farnesyl diphosphate (FPP). These data suggested that, unlike the green peach aphid and the pea aphid, the cotton aphid appears to contain only a single IPPS with dual FPP/GPP synthase activity.
Assuntos
Afídeos/enzimologia , Dimetilaliltranstransferase/genética , Proteínas de Insetos/genética , Animais , Afídeos/genética , Afídeos/crescimento & desenvolvimento , Clonagem Molecular , Dimetilaliltranstransferase/química , Dimetilaliltranstransferase/metabolismo , Dosagem de Genes , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Cinética , Larva/enzimologia , Larva/genética , Larva/crescimento & desenvolvimento , Filogenia , RNA Mensageiro/metabolismo , Análise de Sequência de DNA , Análise de Sequência de Proteína , Homologia Estrutural de ProteínaRESUMO
Wolbachia is a common and widespread group of bacteria found in reproductive tissues of arthropods. These bacteria are transmitted through the cytoplasm of eggs and have evolved various mechanisms for manipulating reproduction of their hosts. Because of important relationship with some major biological issues and implication of biological control and genetic engineering, the study of Wolbachia has under gone an explosive growth in recent years. A tremendous progress has been made based on the advancing molecular diagnostic technique of Wolbachia. In this paper, those molecular diagnostic techniques are detailed and it is aimed to be the primer for the research of Wolbachia in China.