The invention discloses a T7-RNA polymerase mutant. The mutant is obtained by replacing serine at the 43rd site from the N end of an amino acid sequence of wild T7-RNA polymerase with A-type amino acid or B-type amino acid, wherein the A amino acid is tyrosine, phenylalanine, leucine, lysine or aspartic acid, and the B amino acid is tryptophan, isoleucine, arginine, asparagine, glutamine, glutamic acid or proline. The T7-RNA polymerase mutant is suitable for synthesis of RNA internally containing a termination signal and RNA with a hairpin structure formed at the tail end. The method can be widely applied to the aspects of in-vitro transcription, RNA synthesis, gene editing, RNA drug synthesis, in-vivo protein expression or cell-free protein expression in-vitro translation systems, transcription terminator research, RNA-dependent RNA polymerase activity research, biological transcription regulation element synthesis and the like.