Identification of Protein-Phosphate Contacts in the R17 Coat Protein Binding Site

Small RNAs of defined length and sequence can be synthesized enzymatically using T7 RNA polymerase and synthetic DNA templates which contain the T7 promoter. The templates are base paired from $-17$ to +1 and contain a long 5$\sp\prime$ overhang. The products of the transcription reactions have the length and sequence expected for a runoff transcript but may have one or more extra nucleotide at the 3$\sp\prime$ terminus. In addition to the full length products, large amounts of smaller products in the range from 2 to 6 nucleotides are produced. These products appear to be the result of abortive initiation events. Variants in the promoter region from +1 to +6 have been made to increase the variety of RNAs which can be synthesized and their efficiency as templates determined. Finally, the reaction conditions have been optimized to allow for the synthesis of milligram amounts of virtually any RNA from 15 to 35 nucleotides in length.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Milligan, John Fairbanks

• Chemistry, Biochemistry

Identifier

(UMI)AAI8908781

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/70577