45 mutations (insertion, deletion and base substitution) of the G4 Goric were tested for their functional activity in M13 and R199 in vivo. The critical mutants were also assayed for their ability to synthesize pRNA in vitro using SSB and primase. The results demonstrate that the secondary structure and spacing of stem-loops I and III are essential for Goric activity and that the 5′-CTG-3′ sequence flanking stem-loop I is essential for initiation of pRNA synthesis.
Bibliographical noteFunding Information:
We thank Dr. Nobuo Nomura for his communication and gifts of E. coli and phage strains. We also thank Drs. Hisao Masai, Kenneth Marians, Kinya Yoda and Tuneko Okazaki for their valuable discussions and communications prior to publication and Dr. Roger McMacken for providing us the purified primase.Ill This work was supported in part by Public Health Service Grant GM38292 to G.N.G.