Caton A J, Robertson J S
Nucleic Acids Res. 1979 Nov 24;7(6):1445-56. doi: 10.1093/nar/7.6.1445.
A novel technique is described for the production of pure, full-length influenza virus ds DNA's corresponding to each segment of the influenza virus genome, and suitable for molecular cloning and restriction endonuclease mapping. The method involves the synthesis of DNA complementary to both virion (negative strand) and messenger (positive strand) RNA, gel purification and annealing. By avoiding the use of SI nuclease, which often removes the terminal regions of DNA duplexes, the method allows transcription of the total sequence information of influenza virion and messenger RNA's into a ds DNA form.
本文描述了一种新技术,用于生产与流感病毒基因组各片段相对应的纯全长流感病毒双链DNA,适用于分子克隆和限制性内切酶图谱分析。该方法包括合成与病毒粒子(负链)和信使(正链)RNA互补的DNA、凝胶纯化和退火。通过避免使用通常会去除DNA双链末端区域的S1核酸酶,该方法能够将流感病毒粒子和信使RNA的全部序列信息转录成双链DNA形式。