Author： Sousa R.
Publisher： Oxford University Press
ISSN： 1362-4962
Source： Nucleic Acids Research, Vol.30, Iss.24, 2002-12, pp. : 138-138
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Abstract
A T7 RNA polymerase in which Tyr639 is mutated to Phe readily utilizes 2′-deoxy, 2′-NH2 and 2′-F NTPs as substrates and has been widely used to synthesize modified RNAs for a variety of applications. This mutant does not readily utilize NTPs with bulkier 2′-substituents, nor does it facilitate incorporation of NTPs with modifications at other positions. Introduction of a second mutation (H784A) into the Y639F background markedly enhances utilization of NTPs with bulky 2′-substituents (2′-OMe and 2′-N3), and may also enhance use of NTPs with modifications at other than the 2′-position. The Y639F/H784A double mutant may therefore be exceptionally useful for incorporation of a variety of non-canonical NMPs into RNA.
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