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利用杆状病毒载体过表达家蚕促前胸腺激素

Overexpression of Bombyx mori prothoracicotropic hormone using baculovirus vectors.

作者信息

O'Reilly D R, Kelly T J, Masler E P, Thyagaraja B S, Robson R M, Shaw T C, Miller L K

机构信息

Department of Entomology, University of Georgia, Athens 30602, USA.

出版信息

Insect Biochem Mol Biol. 1995 Apr;25(4):475-85. doi: 10.1016/0965-1748(94)00087-f.

Abstract

Recombinant baculoviruses were constructed that express the cDNA encoding the prothoracicotropic hormone (PTTH) of Bombyx mori. This hormone stimulates the production of ecdysteroids by the insect's prothoracic glands. Two groups of viruses were constructed, expressing either the entire cDNA encoding prepro-PTTH, or a synthetic chimeric gene encoding a signal peptide fused to the mature PTTH subunit. In both cases, the genes were expressed in wild-type Autographa californica nuclear polyhedrosis virus (AcMNPV) and in vEGTDEL, an ACMNPV mutant that lacks a functional egt gene. The egt gene is required for viral-mediated inactivation of host ecdysteroids. High levels of functional PTTH were produced only by viruses expressing the mature subunit cDNA. This recombinant PTTH resembled the native hormone by all criteria examined. The overproduction of B. mori PTTH induced higher than normal levels of haemolymph ecdysteroids but had no observable effects on the development of infected Spodoptera frugiperda larvae. However, expression of PTTH by AcMNPV was found to inhibit the pathogenicity of the virus. This effect was particularly marked in the case of viruses lacking a functional egt gene.

摘要

构建了表达家蚕促前胸腺激素(PTTH)编码cDNA的重组杆状病毒。这种激素刺激昆虫前胸腺产生蜕皮甾类。构建了两组病毒,一组表达编码前激素原PTTH的完整cDNA,另一组表达编码与成熟PTTH亚基融合的信号肽的合成嵌合基因。在这两种情况下,基因均在野生型苜蓿银纹夜蛾核型多角体病毒(AcMNPV)和vEGTDEL(一种缺乏功能性egt基因的AcMNPV突变体)中表达。egt基因是病毒介导的宿主蜕皮甾类失活所必需的。只有表达成熟亚基cDNA的病毒才能产生高水平的功能性PTTH。根据所有检测标准,这种重组PTTH与天然激素相似。家蚕PTTH的过量产生诱导血淋巴蜕皮甾类水平高于正常,但对感染的草地贪夜蛾幼虫的发育没有可观察到的影响。然而,发现AcMNPV表达PTTH会抑制病毒的致病性。在缺乏功能性egt基因的病毒中,这种效应尤为明显。

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