Carstens E B, Lu A L, Chan H L
Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada.
J Virol. 1993 May;67(5):2513-20. doi: 10.1128/JVI.67.5.2513-2520.1993.
A 2.8-kb region of the Autographa californica nuclear polyhedrosis virus genome was sequenced and found to contain an open reading frame (p47) which was capable of rescuing a previously characterized temperature-sensitive mutant, ts317 (S. Partington, H. Yu, A. Lu, and E. B. Carstens, Virology 157:91-102, 1990). Transcriptional mapping demonstrated that an early 4.2-kb RNA encoded the p47 open reading frame and probably overlapped the 39K delayed-early gene. The p47 open reading frame was cloned behind the polyhedrin promoter in a baculovirus transfer plasmid, which was then used to prepare a recombinant baculovirus overexpressing the p47 polypeptide. The overexpressed polypeptide was used to prepare p47-specific monoclonal antibodies. These antibodies detected a polypeptide of 47 kDa in A. californica nuclear polyhedrosis virus-infected cells, demonstrating that p47 is expressed as an authentic viral product. The p47 gene product was localized to the nucleus of infected cells, supporting the hypothesis that it is involved in regulating viral transcription at late times postinfection.
对苜蓿银纹夜蛾核型多角体病毒基因组的一个2.8kb区域进行了测序，发现其中包含一个开放阅读框（p47），该阅读框能够拯救一个先前已鉴定的温度敏感突变体ts317（S. Partington、H. Yu、A. Lu和E. B. Carstens，《病毒学》157:91 - 102，1990年）。转录图谱分析表明，一个4.2kb的早期RNA编码p47开放阅读框，并且可能与39K延迟早期基因重叠。将p47开放阅读框克隆到杆状病毒转移质粒的多角体蛋白启动子之后，然后用其制备过表达p47多肽的重组杆状病毒。用过表达的多肽制备p47特异性单克隆抗体。这些抗体在苜蓿银纹夜蛾核型多角体病毒感染的细胞中检测到一条47kDa的多肽，表明p47作为一种真实的病毒产物被表达。p47基因产物定位于感染细胞的细胞核，支持了它在感染后期参与调节病毒转录的假说。
