Morgeaux S, Tordo N, Gontier C, Perrin P
WHO Collaborating Centre for Reference and Research on Rabies, Institut Pasteur, Paris, France.
Vaccine. 1993;11(1):82-90. doi: 10.1016/0264-410x(93)90343-v.
The effects of beta-propiolactone (BPL), an alkylating and virus inactivating agent, on the structural and in vitro biological properties of different DNA preparations from BHK-21 cells were investigated. Both uninfected and rabies virus-infected cells were used. Purified cellular DNA (celDNA) was used as the reference, and supernatants from infected cells were treated with BPL. For structural and biological studies three types of DNA preparation were tested: celDNA; purified DNA from cell (infected or uninfected) supernatant (pcsDNA) with or without BPL treatment; and residual cell DNA present in purified rabies virus (inactivated or not) preparations. Rabies infection and BPL (diluted 1:4000) treatment induced modifications in the structure of the three DNA types, including strand breaks and nicks. The damage to the DNA structure by BPL modifies the biological properties of the pcsDNA appraised by its ability to serve as the template in vitro for different polymerases. When rabies virus was inactivated with BPL diluted 1:1000 the DNA damage increased dramatically: small double-stranded DNA fragments (50-200 base pairs) were generated which could not function as templates for polymerases.
研究了烷基化和病毒灭活剂β-丙内酯（BPL）对来自BHK-21细胞的不同DNA制剂的结构和体外生物学特性的影响。使用了未感染和感染狂犬病病毒的细胞。以纯化的细胞DNA（celDNA）作为对照，并用BPL处理感染细胞的上清液。为了进行结构和生物学研究，测试了三种类型的DNA制剂：celDNA；来自细胞（感染或未感染）上清液的纯化DNA（pcsDNA），有或没有BPL处理；以及纯化的狂犬病病毒（灭活或未灭活）制剂中存在的残留细胞DNA。狂犬病感染和BPL（稀释1:4000）处理导致三种DNA类型的结构发生改变，包括链断裂和切口。BPL对DNA结构的损伤改变了pcsDNA的生物学特性，通过其作为不同聚合酶体外模板的能力来评估。当用稀释1:1000的BPL灭活狂犬病病毒时，DNA损伤急剧增加：产生了小的双链DNA片段（50-200个碱基对），这些片段不能作为聚合酶的模板。
