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粉纹夜蛾颗粒体病毒病毒增强因子编码基因的定位与核苷酸序列

Location and nucleotide sequence of the gene encoding the viral enhancing factor of the Trichoplusia ni granulosis virus.

作者信息

Hashimoto Y, Corsaro B G, Granados R R

机构信息

Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, New York 14853-1801.

出版信息

J Gen Virol. 1991 Nov;72 ( Pt 11):2645-51. doi: 10.1099/0022-1317-72-11-2645.

Abstract

The gene encoding the viral enhancing factor (VEF) of Trichoplusia ni granulosis virus has been cloned from a lambda gt11 expression library, and the complete nucleotide sequence determined. The VEF gene encodes a protein with a predicted Mr of 104K which does not share homology with any previously reported proteins. A possible promoter is located four nucleotides upstream of the initiation codon and represents a consensus baculovirus late promoter (ATAAG). This has been confirmed by the identification of VEF mRNA in Northern blots of infected larvae 6 days but not 3 days post-infection. Using an anti-VEF-TrpE polyclonal antiserum in Western blots of dissolved viral occlusion bodies, related proteins have been identified in both Pseudaletia unipuncta granulosis virus Hawaiian strain (PuGV-H) and Heliothis armigera GV (HaGV), but not in Erinnyis ello GV (EeGV), T. ni singly enveloped nuclear polyhedrosis virus (TnSNPV) or Autographa californica multiply enveloped NPV (AcMNPV). Similar results were obtained with Southern blots of genomic digests. DNA fragments homologous to an internal portion of the VEF gene were found in PuGV-H and HaGV but not in EeGV, TnSNPV or AcMNPV.

摘要

粉纹夜蛾颗粒体病毒的病毒增强因子(VEF)编码基因已从λgt11表达文库中克隆出来,并测定了其完整的核苷酸序列。VEF基因编码一种预测分子量为104K的蛋白质,该蛋白质与任何先前报道的蛋白质均无同源性。一个可能的启动子位于起始密码子上游四个核苷酸处,代表杆状病毒晚期启动子共有序列(ATAAG)。这已通过在感染后6天而非3天的受感染幼虫的Northern印迹中鉴定VEF mRNA得到证实。在溶解的病毒包涵体的Western印迹中使用抗VEF-TrpE多克隆抗血清,在草地贪夜蛾颗粒体病毒夏威夷株(PuGV-H)和棉铃虫GV(HaGV)中均鉴定出相关蛋白,但在眉纹天蚕蛾GV(EeGV)、粉纹夜蛾单囊核型多角体病毒(TnSNPV)或苜蓿银纹夜蛾多囊核型多角体病毒(AcMNPV)中未鉴定出。用基因组消化产物的Southern印迹也得到了类似结果。在PuGV-H和HaGV中发现了与VEF基因内部部分同源的DNA片段,但在EeGV、TnSNPV或AcMNPV中未发现。

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