Winter G, Fields S
Nucleic Acids Res. 1980 May 10;8(9):1965-74. doi: 10.1093/nar/8.9.1965.
A strategy has been developed for sequencing the single-stranded RNA genes of influenza virus. Restriction fragments derived from double-stranded cDNA copies of total influenza RNA were cloned into the bacteriophage M13mp2 and sequenced by the dideoxy technique. Sequences were extended and overlapped by using the virion RNA as template and priming with small restriction fragments. In the course of this strategy, the nucleotide sequence of segment 7 (1027 nucleotides) was completed and provides the primary structure of the matrix protein (27, 861 daltons). In addition, there is a second long reading frame which partly overlaps the reading frame of the matrix protein.
已开发出一种用于对流感病毒单链RNA基因进行测序的策略。从流感病毒总RNA的双链cDNA拷贝中获得的限制性片段被克隆到噬菌体M13mp2中，并通过双脱氧技术进行测序。以病毒粒子RNA为模板，用小的限制性片段进行引物延伸，使序列得以扩展并重叠。在该策略实施过程中，完成了第7节段（1027个核苷酸）的核苷酸序列测定，该序列提供了基质蛋白（27861道尔顿）的一级结构。此外，还有第二个长阅读框，它与基质蛋白的阅读框部分重叠。
