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位于苜蓿银纹夜蛾核型多角体病毒多角体蛋白基因下游的1629个核苷酸的开放阅读框编码一种与核衣壳相关的磷蛋白。

The 1,629-nucleotide open reading frame located downstream of the Autographa californica nuclear polyhedrosis virus polyhedrin gene encodes a nucleocapsid-associated phosphoprotein.

作者信息

Vialard J E, Richardson C D

机构信息

Department of Microbiology and Immunology, McGill University, Montréal, Québec, Canada.

出版信息

J Virol. 1993 Oct;67(10):5859-66. doi: 10.1128/JVI.67.10.5859-5866.1993.

Abstract

A 78-kDa protein was produced in bacteria from a clone of the 1,629-nucleotide open reading frame located immediately downstream from the polyhedrin gene of Autographa californica nuclear polyhedrosis virus. The identity of this protein was confirmed by its reactivity with peptide antiserum and amino terminal peptide sequencing after purification from transformed bacteria. The polypeptide was used to produce polyclonal antisera in rabbits. Immunoblot analysis of insect cells infected with the baculovirus indicated that two related proteins with molecular masses of 78 and 83 kDa were synthesized late in infection. Biochemical fractionation studies indicated that both of these proteins were present in purified nucleocapsids from budded and occluded virus preparations. Immunoprecipitation of 32P-labeled proteins and treatment of purified nucleocapsids with alkaline phosphatase demonstrated that the 83-kDa protein was a phosphorylated derivative of the 78-kDa protein. Furthermore, immunoelectron microscopy revealed that the proteins were localized to regions of nucleocapsid assembly within the infected cell and appeared to be associated with the end structures of mature nucleocapsids.

摘要

一种78 kDa的蛋白质是由细菌从一个1629个核苷酸的开放阅读框克隆体中产生的,该开放阅读框位于苜蓿银纹夜蛾核型多角体病毒多角体蛋白基因的紧下游。从转化细菌中纯化后,通过其与肽抗血清的反应性和氨基末端肽测序确认了该蛋白质的身份。该多肽用于在兔中产生多克隆抗血清。对感染杆状病毒的昆虫细胞进行免疫印迹分析表明,感染后期合成了两种分子量分别为78 kDa和83 kDa的相关蛋白质。生化分级分离研究表明,这两种蛋白质都存在于从出芽病毒制剂和包涵体病毒制剂中纯化的核衣壳中。对32P标记蛋白质的免疫沉淀以及用碱性磷酸酶处理纯化的核衣壳表明,83 kDa的蛋白质是78 kDa蛋白质的磷酸化衍生物。此外,免疫电子显微镜显示,这些蛋白质定位于感染细胞内核衣壳组装区域,并且似乎与成熟核衣壳的末端结构相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed1/238004/2794fd0c47ec/jvirol00031-0163-a.jpg

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