New method for isolation and sequence determination of 5'-terminal regions of bacteriophage phiX174 in vitro mRNAs.
We have determined the nucleotide sequences of the 5'-terminal oligonucleotides, produced by RNase T1 digestion of bacteriophage phiX174 mRNAs synthesized in vitro. The major sequences are: pppCpGp(Ap), pppApUpCpGp(Cp), pppAp(Ap)2UpCp(Up)2Gp(Gp), and pppAp(Ap)3UpCp(Up)2Gp(Gp). The sequences of several minor 5'-terminal oligonucleotides were also determined. For this research we have devised a simple isolation procedure, for the 5'-terminal oligonucleotides, based upon hydroxylapatite chromatography and two-dimensional thin-layer separation. This method allows for the rapid and quantitative recovery of all oligonucleotides containing 5'-triphosphate end groups and should be generally useful for sequence on 5' termini of mRNAs.
D001483:Base Sequence; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D003090:Coliphages / Q000032:analysis*; D004267:DNA Viruses / Q000032:analysis; D006882:Hydroxyapatites; D008722:Methods; D009841:Oligonucleotides / Q000032:analysis; D012333:RNA, Messenger / Q000302:isolation & purification; D012367:RNA, Viral / Q000302:isolation & purification; D012260:Ribonucleases; D012265:Ribonucleotides / Q000032:analysis
D006882:Hydroxyapatites; D009841:Oligonucleotides; D012333:RNA, Messenger; D012367:RNA, Viral; D012265:Ribonucleotides; D012260:Ribonucleases