Superior 5' homogeneity of RNA from ATP-initiated transcription under the T7 phi 2.5 promoter.
Transcription from the commonly used GTP- initiating T7 class III promoter phi6.5 frequently produces heterogeneous RNA at both 3' and 5' ends. We demonstrate here that RNA transcripts from the T7 class II promoter phi2.5 have superior 5' homogeneity over those from the phi6.5 promoter, with comparable total RNA yields. The overall homogeneity of RNA transcripts is improved to different degrees depending on RNA sequences, although transcription under phi2.5 does not affect the 3' heterogeneity of RNA. In combination with 3' RNA trimming by DNAzymes or ribozymes, this ATP- initiated transcription system based on the T7 phi2.5 promoter can provide excellent quality of RNA for applications requiring a high degree of RNA size homogeneity.
Coleman
Tricia M
TM
Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406-5043, USA.
Wang
Guocan
G
Huang
Faqing
F
D020413:3' Untranslated Regions / Q000235:genetics / Q000378:metabolism; D020121:5' Untranslated Regions / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism* / Q000494:pharmacology; D017123:Bacteriophage T7 / Q000235:genetics*; D001483:Base Sequence; D015967:Gene Expression Regulation, Viral; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D012367:RNA, Viral; D000255:Adenosine Triphosphate