水痘-带状疱疹病毒基因组作为感染性细菌人工染色体在大肠杆菌中的克隆。

Cloning of the varicella-zoster virus genome as an infectious bacterial artificial chromosome in Escherichia coli.

作者信息

Nagaike Kazuhiro, Mori Yasuko, Gomi Yasuyuki, Yoshii Hironori, Takahashi Michiaki, Wagner Markus, Koszinowski Ulrich, Yamanishi Koichi

机构信息

Department of Microbiology, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka 565-0871, Japan.

出版信息

Vaccine. 2004 Sep 28;22(29-30):4069-74. doi: 10.1016/j.vaccine.2004.03.062.

DOI:10.1016/j.vaccine.2004.03.062

PMID:15364458

Abstract

The complete genome of the varicella-zoster virus (VZV) Oka strain has been cloned as a bacterial artificial chromosome (BAC). Following electroporation into Escherichia coli (E. coli) strain DH10B, the VZV BAC was stably propagated over multiple generations of its host. Human embryonic lung (HEL) cells transfected with VZV BAC DNA recovered from DH10B showed cytopathic effect (CPE), and virus spread to neighbouring cells was observed. BAC vector sequences are flanked by loxP sites and, coinfection of the reconstituted virus, with a recombinant adenovirus expressing Cre recombinase removed the bacterial sequences. The resulting recombinant rV02 grew as well as the parental virus in HEL cells. The recombinant VZV will promote VZV research and increase use of the viral genome as an investigative tool.

摘要

水痘-带状疱疹病毒（VZV）Oka株的全基因组已被克隆为细菌人工染色体（BAC）。将其电穿孔导入大肠杆菌DH10B菌株后，VZV BAC在其宿主的多代培养中稳定繁殖。用从DH10B中回收的VZV BAC DNA转染人胚肺（HEL）细胞，出现细胞病变效应（CPE），并观察到病毒扩散至邻近细胞。BAC载体序列两侧为loxP位点，重组病毒与表达Cre重组酶的重组腺病毒共感染可去除细菌序列。所得重组体rV02在HEL细胞中的生长情况与亲本病毒相同。重组VZV将促进VZV研究，并增加病毒基因组作为研究工具的应用。