REPLICATION OF ORGYIA-PSEUDOTSUGATA BACULOVIRUS DNA - LEF-2 AND IE-1 ARE ESSENTIAL AND IE-2, P34, AND OP-IAP ARE STIMULATORY GENES
REPLICATION OF ORGYIA-PSEUDOTSUGATA BACULOVIRUS DNA - LEF-2 AND IE-1 ARE ESSENTIAL AND IE-2, P34, AND OP-IAP ARE STIMULATORY GENES
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DOI：
10.1006/viro.1995.1523
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发表时间：
1995-10-01
期刊：
影响因子：
3.7
通讯作者：
ROHRMANN, GF
中科院分区：
文献类型：
--
作者：
AHRENS, CH;ROHRMANN, GF
瞬时DNA复制试验用于鉴定位于微囊藻毒素中的基因。假糖纹夜蛾多核衣壳核型多角体病毒(OpMNPV)基因组的90.5-7.0，当与未感染的Lymantria disdisar细胞共感染时，影响含有OpMNPV复制起点的报告质粒的复制。病毒反式激活因子ie-1和一个2.4kb的亚克隆被发现是复制所必需的。对2.4kb的区域进行了测序，并确定了开放阅读框。利用该区域的亚克隆进行复制分析，确定了一个名为晚期表达因子2(lef-2)的基因是必需的复制基因。OpMNPV lef-2基因编码一个分子量为22.7 kDa(204个氨基酸)的蛋白，其氨基酸序列与其同源序列有54.7%的同源性。Northern杂交和S1核酸酶保护分析的转录图谱表明，OpMNPV lef-2在感染后的早期和晚期都以约1.6kb的转录本的形式表达。早期转录起始于TAATA盒下游约30个核苷酸，而晚期转录起始于晚期启动子一致基序内。此外，我们还鉴定了三个刺激DNA复制的基因，包括两个OpMNPV转录激活子(ie-2和p34)和Op-IAP，Op-IAP是AcMNPV p35基因的功能类似物，它抑制了感染AcMNPV的果蝇细胞的凋亡。(C)1995年学术出版社。
A transient DNA replication assay was used to identify genes located within m.u. 90.5-7.0 of the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (OpMNPV) genome that influenced replication of a reporter plasmid containing an OpMNPV origin of replication, when cotransfected into uninfected Lymantria dispar cells. The viral transactivator ie-1 and a 2.4-kb subclone were found to be essential for replication. The 2.4-kb region was sequenced and open reading frames were identified. Replication assays using subclones from this region identified a gene called late expression factor 2 (lef-2), as the essential replication gene. The OpMNPV lef-2 gene encodes a protein with a predicted molecular weight of 22.7 kDa (204 amino acids) and exhibits 54.7% amino acid sequence identity with its homolog in the genome of the Autographa californica MNPV. Transcriptional mapping using both Northern blot and S1 nuclease protection assays demonstrated that OpMNPV lef-2 was expressed at both early and late times postinfection as a transcript of about 1.6 kb. The early transcript initiated approximately 30 nt downstream of a TAATA box, whereas the late transcript initiated from within a late promoter consensus motif. In addition, we identified three genes stimulatory for DNA replication including two OpMNPV transcriptional activators (ie-2 and p34) and Op-iap, which is the functional analog of the AcMNPV p35 gene that inhibits apoptosis in AcMNPV-infected Spodoptera frugiperda cells. (C) 1995 Academic Press, Inc.