T4 RNA ligase 2 mutants and preparation method and use thereof

Abstract

Provided are T4 RNA ligase 2 mutants and preparation method and use thereof, relating to the field of biology and enzyme engineering. The T4 ligase 2 mutants have mutation at least one site of positions 62, 103, 166, 168, 193, 217, 260, 297, 298, 303, 306, 311, 313 and 318 of sequence of the wild-type T4 RNA ligase 2. Compared with the wild type, the mutants have remarkably improved thermal stability, and can maintain relatively high ligase activity at higher temperatures such as 45° C., and 50° C. In addition to ligating natural nucleic acid sequences, the mutants further can ligate non-natural nucleic acids with modification at high temperatures. The mutants can synthesize ribonucleic acids at elevated temperatures, so as to effectively reduce the production of byproducts caused by base mismatching, thus improving the efficiency of RNA synthesis, and facilitating large-scale production of high-quality RNAs.

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