通过SV40转录和复制信号，从克隆的cDNA产生感染性脊髓灰质炎病毒的效率显著提高。
Production of infectious poliovirus from cloned cDNA is dramatically increased by SV40 transcription and replication signals.
作者信息
Semler B L, Dorner A J, Wimmer E
出版信息
Nucleic Acids Res. 1984 Jun 25;12(12):5123-41. doi: 10.1093/nar/12.12.5123.
Abstract
Sub-genomic cDNA clones representing the entire genomic RNA of poliovirus Type 1 (Mahoney) have been isolated in E. coli. Construction of a complete cDNA copy of the poliovirus genome in the EcoRI site of plasmid vector pBR325 from these clones is described. Introduction of plasmid DNA containing the complete cDNA copy of polio RNA into cultured primate cells by transfection produces infectious poliovirus. The virus produced by such a transfection appears to be identical to wild type poliovirus. Isolation of a polio recombinant plasmid containing SV40 transcription and replication signals is also described. Transfection of COS-1 cells with this plasmid yields greater than 1,600 plaque-forming units (PFU) per microgram of input DNA.
摘要
代表1型脊髓灰质炎病毒（Mahoney株）全基因组RNA的亚基因组cDNA克隆已在大肠杆菌中分离出来。本文描述了从这些克隆中在质粒载体pBR325的EcoRI位点构建脊髓灰质炎病毒基因组的完整cDNA拷贝。通过转染将含有脊髓灰质炎病毒RNA完整cDNA拷贝的质粒DNA导入培养的灵长类细胞中可产生感染性脊髓灰质炎病毒。这种转染产生的病毒似乎与野生型脊髓灰质炎病毒相同。本文还描述了含有SV40转录和复制信号的脊髓灰质炎重组质粒的分离。用该质粒转染COS-1细胞，每微克输入DNA可产生超过1600个空斑形成单位（PFU）。
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