MRNA Cap2'O methyltransferase mRNA Cap2'O methyltransferase is derived from a recombinant Escherichia coli strain carrying the cowpox virus mRNA Cap2'O methyltransferase gene. This enzyme can add a methyl group to the 2'O position of the first nucleotide at the 5'end of RNA, adjacent to the cap structure. S-adenosylmethionine SAM can also be used as a methyl donor to methylate the capped RNA Cap 0). The Cap1 structure can enhance the translation efficiency of mRNA, thus helping to improve mRNA expression in transfection and microinjection experiments. This enzyme requires RNA with a m7GpppN or 7-methylguanosine cap structure as the substrate, and RNA with pN ppN pppN or GpppN at the 5 'end cannot be used. Capped RNA can be prepared by in vitro transcription using cap like analogs or by enzymatic reaction using Vaccinia Capping Enzymes.
Product storage -2 5 1515 ℃ storage (to avoid repeated freeze-thaw)
Storage buffer: 20 mM Tris HCl pH 8.0 (25 ℃ 25 ℃), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.1% Triton X 100 50% glycol
Activity definition: The amount of enzyme required to methylate 10 pmol of 80nt capped RNA transcripts within 1 hour at 37 ℃ is defined as one active unit
Exonuclease activity: 50 U of this product and 1 μ G λ Hind III digest DNA reacted at 37 ℃ for 16 hours without any changes in the electrophoretic bands of the DNA;
Endonuclease activity: 50 U of this product and 1 μ G λ The electrophoresis bands of DNA did not change after reacting at 37 ℃ for 16 hours;
Incision enzyme activity: 50 U of this product and 1 μ The electrophoretic bands of DNA did not change when g pBR 322 reacted at 37 ℃ for 16 hours;
RNase activity: 50 U of this product and 1.6 μ The electrophoresis bands of MS2 RNA did not change after reacting at 37 ℃ for 4 hours;
Escherichia coli DNA residue detection: The residual nucleic acid in 50 U of this product was detected by E. coli 16s rDNA specific Taq Man qPCR, and the E. coli genome residue was ≤ 1 copy.
Bacterial endotoxin (Endotoxin LAL Test, General Rule of gel Limit Test, Vol. 4, Chinese Pharmacopoeia 2020, 1143), bacterial endotoxin content ≤ 10 EU/mg.