Cloning of the gene encoding the antibacterial peptide drosocin involved in Drosophila immunity - Expression studies during the immune response
Cloning of the gene encoding the antibacterial peptide drosocin involved in Drosophila immunity - Expression studies during the immune response
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DOI：
10.1111/j.1432-1033.1996.00699.x
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发表时间：
1996-11-01
期刊：
影响因子：
--
通讯作者：
Meister, M
中科院分区：
文献类型：
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作者：
Charlet, M;Lagueux, M;Meister, M
最近从果蝇[Bulet，P.，Dimarcq，J.L.，Hetru，C.，Lagueux，M.，Charlet，M.，Hegy，G.，Van Dorsselaer，A.和Hoffman，J.A.(1993)J.Biol]中分离到一种携带O-糖基化取代基的有效可诱导抗菌肽。化学。268,14893-14897]在这里，我们报告的克隆研究表明，果蝇包含一个单一的。无内含子基因，位于51C1-6位，编码用于加工Droocin的前体蛋白。Droocin基因的上游和下游序列含有与哺乳动物调控基序相似的顺式调控元件，即三个与kappa k-B相关的十聚体序列。在幼年动物中，Droocin基因是沉默的，在幼虫和成虫中免疫攻击后，Droocin基因以急性期动力学强烈诱导。我们已经建立了几个报告基因被置于不同Droocin启动子序列控制下的转基因果蝇品系。我们的结果表明，2.5kb的上游序列对转基因具有诱导性和组织特异性，但其在免疫攻击后在脂肪体中的表达水平较低。在Droocin转录序列下游增加基因组区域会导致转录水平的提高，这与感染时融合和驻留的Droocin基因相似。对转基因果蝇品系的分析表明，Droocin报告基因在产卵雌性的输卵管中结构性表达。
A potent inducible antibacterial peptide carrying an O-glycosylated substitution has recently been isolated from Drosophila [Bulet, P., Dimarcq, J. L., Hetru, C., Lagueux, M., Charlet, M., Hegy, G., Van Dorsselaer, A. and Hoffman, J. A. (1993) J. Biol. Chem. 268, 14893-14897]. Here we report cloning studies that show that Drosophila contains a single. intronless gene, located at position 51C1-6, which encodes the precursor protein from which drosocin is processed. The upstream and the downstream sequences of the drosocin gene contain putative cis-regulatory elements similar to mammalian regulatory motifs, namely three kappa k-B-related decameric sequences. The drosocin gene is silent in naive animals, and is strongly induced with acute phase kinetics after immune challenge in larvae and in adults. We have established several transgenic fly lines in which reporter genes were placed under the control of various drosocin promoter sequences. Our results indicate that 2.5 kb of upstream sequences confer inducibility and tissue specificity to the transgene, but that the level of its expression in the fat body after immune challenge is low. Addition of genomic regions downstream of the drosocin transcribed sequences results in increased transcription levels, which are similar for the fusion and the resident drosocin genes upon infection. Analysis of transgenic fly lines showed that the drosocin reporter gene is constitutively expressed in the oviducts of egg-laying females.