Hauser C, Fusswinkel H, Li J, Oellig C, Kunze R, Müller-Neumann M, Heinlein M, Starlinger P, Doerfler W
Institute of Genetics, University of Cologne, Federal Republic of Germany.
Mol Gen Genet. 1988 Nov;214(3):373-8. doi: 10.1007/BF00330469.
The polypeptide encoded in the Activator (Ac) element of Zea mays L. has been expressed in Spodoptera frugiperda insect cells using plasmids which carry the strong polyhedrin promoter of the baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV). Recombinant AcNPVs with the Ac-cDNA integrated and under the control of the viral polyhedrin promoter have been isolated and their genomes have been partly characterized as to the location of the foreign DNA insert. Upon infection of S. frugiperda cells with the recombinant AcNPV, maize Ac element specific messenger RNAs, as well as a newly synthesized polypeptide with an apparent molecular weight of about 116 kDa, have been detected in extracts of recombinant infected cells. This polypeptide is absent from extracts of wild-type infected cells expressing the polyhedrin polypeptide which can be recognized by the presence of nuclear inclusion bodies. Recombinant infected cells lack this protein. The Ac specific polypeptide is detected by antisera, which have been raised against fusion proteins containing Ac sequences synthesized in Escherichia coli, both in immunoprecipitation and in Western blotting experiments. The Ac specific protein is a nuclear phosphoprotein and represents about 1%-2% of the newly synthesized protein.
利用携带苜蓿银纹夜蛾核型多角体病毒（AcNPV）强多角体蛋白启动子的质粒，在草地贪夜蛾昆虫细胞中表达了玉米（Zea mays L.）激活子（Ac）元件编码的多肽。已分离出整合了Ac - cDNA并受病毒多角体蛋白启动子控制的重组AcNPV，并对其基因组中外源DNA插入片段的位置进行了部分鉴定。用重组AcNPV感染草地贪夜蛾细胞后，在重组感染细胞的提取物中检测到了玉米Ac元件特异性信使RNA，以及一种新合成的表观分子量约为116 kDa的多肽。在表达可通过核内含体识别的多角体蛋白多肽的野生型感染细胞提取物中不存在这种多肽。重组感染细胞缺乏这种蛋白质。在免疫沉淀和蛋白质印迹实验中，抗血清可检测到Ac特异性多肽，该抗血清是针对含有在大肠杆菌中合成的Ac序列的融合蛋白产生的。Ac特异性蛋白是一种核磷蛋白，约占新合成蛋白的1% - 2%。
