期刊
NUCLEIC ACIDS RESEARCH
卷 43, 期 15, 页码 7480-7488出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkv734
关键词
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资金
- NIH [GM083703, 5U54EB015403-02]
- Welch Foundation [H-1592, F-1654]
- National Security Science and Engineering Faculty Fellowship [FA9550-10-1-0169]
- NSSEFF [FA9550-10-1-0169]
On average, mutations are deleterious to proteins. Mutations conferring new function to a protein often come at the expense of protein folding or stability, reducing overall activity. Over the years, a panel of T7 RNA polymerases have been designed or evolved to accept nucleotides with modified ribose moieties. These modified RNAs have proven useful, especially in vivo, but the transcriptional yields tend to be quite low. Here we show that mutations previously shown to increase the thermal tolerance of T7 RNA polymerase can increase the activity of mutants with expanded substrate range. The resulting polymerase mutants can be used to generate 2'-O-methyl modified RNA with yields much higher than enzymes currently employed.
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