Braunagel S C, He H, Ramamurthy P, Summers M D
Department of Entomology, Texas A&M University, College Station 77843-2475, USA.
Virology. 1996 Aug 1;222(1):100-14. doi: 10.1006/viro.1996.0401.
This paper identifies two structural proteins of the occluded derived viral envelope of Autographa californica nuclear polyhedrosis virus (AcMNPV): ODV-E18 and ODV-E35. In addition, we identify a protein, ODV-EC27, that is incorporated into the capsid of occluded virus, which is not detected in budded virus. The genes for these proteins reside within the IE0 intron. The intron was sequenced, and five open reading frames (ORF) were identified. ORF 3 (genomic ORF 143) codes for the ODV envelope protein, ODV-E18. ORF 4 (genomic ORF 144) codes for ODV-EC27, and Western blot analyses locate this protein to both the ODV capsid and envelope. Transcripts for both ODV-E18 and ODV-EC27 initiate from conserved TAAG motifs, and transcripts are detected from 16 through 72 hr p.i. Antiserum to ODV-E18 recognizes a band of 18 kDa on Western blots of extracts from infected cells and bands of 18 and 35 kDa on Western blots of proteins from purified ODV envelope. N-terminal amino acid sequencing reveals that both ODV-E18 and ODV-E35 contain the same N-terminus. Antiserum to ODV-EC27 recognizes a protein of 27 kDa on Western blots of extracts from infected cells and bands of 27 and 35 kDa on Western blots of proteins from purified ODV. Using immunogold labeling techniques, ODV-E18 and/or ODV-E35 are detected in viral induced intranuclear microvesicles and are not detected in the plasma membrane, cytoplasmic membranes, or the nuclear envelope. Immunogold labeling using antisera to ODV-EC27 detects this protein on both the ODV envelope and capsid.
本文鉴定了苜蓿银纹夜蛾核型多角体病毒(AcMNPV)包涵体衍生病毒包膜的两种结构蛋白:ODV-E18和ODV-E35。此外,我们鉴定出一种蛋白ODV-EC27,它被整合到包涵体病毒的衣壳中,而在出芽病毒中未检测到。这些蛋白的基因位于IE0内含子内。对该内含子进行了测序,并鉴定出五个开放阅读框(ORF)。ORF 3(基因组ORF 143)编码ODV包膜蛋白ODV-E18。ORF 4(基因组ORF 144)编码ODV-EC27,蛋白质印迹分析将该蛋白定位到ODV衣壳和包膜上。ODV-E18和ODV-EC27的转录本均从保守的TAAG基序起始,在感染后16至72小时可检测到转录本。抗ODV-E18血清在感染细胞提取物的蛋白质印迹上识别出一条18 kDa的条带,在纯化的ODV包膜蛋白的蛋白质印迹上识别出18 kDa和35 kDa的条带。N端氨基酸测序表明,ODV-E18和ODV-E35具有相同的N端。抗ODV-EC27血清在感染细胞提取物的蛋白质印迹上识别出一种27 kDa的蛋白,在纯化的ODV蛋白的蛋白质印迹上识别出27 kDa和35 kDa的条带。使用免疫金标记技术,在病毒诱导的核内微泡中检测到ODV-E18和/或ODV-E35,而在质膜、细胞质膜或核膜中未检测到。使用抗ODV-EC27血清进行免疫金标记,在ODV包膜和衣壳上均检测到该蛋白。