Customized RNA Sequence Analysis

A customized RNA (5′-GCAUCAGAAAUACACCCGUAGGGCUUU-GAGA-3′) with all the dinucleotide combinations (4² = 16) and four homopolymer sequences was purchased from Integrated DNA Technology (IDT). Yeast tRNA^Phe was obtained from Sigma, and all other chemicals were procured from Thermo Fisher Scientific. RNase T1 was obtained from Worthington Biochemical Corporation. RNase MC1 [11 (link)] and cusativin [12 (link)] were purified on a nickel column using His-tag protein purification kit (EMD Millipore) following their expression from recombinant plasmids in Rosetta ^TM (DE3) and BL21 strains of Escherichia coli, as described before.

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Thakur P., Atway J., Limbach P.A, & Addepalli B. (2022). RNA Cleavage Properties of Nucleobase-Specific RNase MC1 and Cusativin Are Determined by the Dinucleotide-Binding Interactions in the Enzyme-Active Site. International Journal of Molecular Sciences, 23(13), 7021.

Publication 2022

Yeast tRNAPhe RNase T1 His-tag protein purification kit

Cusativin Dinucleotide Escherichia coli Nickel Plasmids Protein Rnase Rnase t1 Strains Trnaphe Yeast

Corresponding Organization : University of Cincinnati

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Variable analysis

independent variables

Customized RNA (5′-GCAUCAGAAAUACACCCGUAGGGCUUU-GAGA-3′) with all the dinucleotide combinations (4^2 = 16) and four homopolymer sequences

dependent variables

Not explicitly mentioned

control variables

Yeast tRNA^Phe obtained from Sigma
All other chemicals procured from Thermo Fisher Scientific
RNase T1 obtained from Worthington Biochemical Corporation
RNase MC1 and cusativin purified on a nickel column using His-tag protein purification kit (EMD Millipore) following their expression from recombinant plasmids in Rosetta™ (DE3) and BL21 strains of Escherichia coli

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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