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KMID : 0578319910010020203
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Molecules and Cells
1991 Volume.1 No. 2 p.203 ~ p.209
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Terminator-distal Transcribed Sequences Affecting Factor-independent Termination Efficiency of Phage T7 Terminator
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Lee, Jong Tae
Kim, Hojeung/Moon, Kyungduk/Kim, Sungjoon/Kang, Changwon
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Abstract
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There is one factor-independent transcription terminator, called To, in phage T7 genome for the phage-encoded RNA polymerase. Termination efficiency of this intrinsic terminator of 41 bp, surrounded by various DNA sequences, was determined in in vitro transcription reactions including linearized plasmid templates. Although the intrinsic terminator was always directly flanked by its upstream genomic sequence of 70 bp, its in vitro termination efficiency varied from 31% to 821/o, while experimental error range is 2%. The variation of the efficiency depended only on the transcribed sequences within a transcription unit, while the sequences outside of the unit did not have any effect. In all cases, however, the hairpin RNA structure of the terminator seems to be formed, as predicted by the energy minimization method of RNA folding. Also there seems to be no particular sequences that enhance or reduce the efficiency regardless of their positions, and the affecting sequences are not just localized to initially transcribed region. Thus, any transcribed DNA sequences even distal to a transcription terminator can affect the efficiency of far downstream factor-independent terminator, apparently without disruption of RNA hairpin terminator structure.
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