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coli DH5α was utilized as a plasmid host strain and cultured in Luria-Bertani broth (LB) (Oxoid)

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The plasmid DNA was linearised with the restriction enzyme Spe1 (New England BioLabs, Inc) and transcribed into mRNA by T7 RNA polymerase with the AMBION mMESSAGE mMACHINE TM T7 transcription kit (Invitrogen TM , Thermo Fisher Scientific)