A new approach to determination of the nucleotide sequence of a viral RNA

✍ Scribed by M. A. Billeter; J. E. Dahlberg; H. M. Goodman; J. Hindley; C. Weissmann

• Publisher: John Wiley and Sons
• Year: 1969
• Tongue: English
• Weight: 596 KB
• Volume: 74
• Category: Article
• ISSN: 0021-9541
• DOI: 10.1002/jcp.1040740420

✦ Synopsis

A QB RNA segment was produced i n vitro by means of a synchronized minus-strand-directed synthesis using purified QP replicase, a reaction known to yield infectious QB RNA. Arguments are presented that a 5.5s product obtained by incubation for 30 seconds at 2 0 °C corresponds to the 5'-terminal region of Q B RNA. After obtaining and sequencing 26 T1 and 21 pancreatic RNase products, we established 3' nearest neighbors by using products labeled with only one nucleoside Ce32Pltriphosphate at a time. The ordering of the oligonucleotides was facilitated by observing the time of their appearance i n products from incubations of 5, 10, 15, 20, and 25 seconds. The enzymatic product consists of 160 nucleotides; no known initiation triplet appears before the sixtieth nucleotide. There is no indication that the coat protein cistron starts within the RNA segment analyzed.