A New Method to Characterize the Translation Initiation Sites of Messenger Ribonucleic Acids
Pease, James Roger
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https://hdl.handle.net/2142/70508
Description
Title
A New Method to Characterize the Translation Initiation Sites of Messenger Ribonucleic Acids
Author(s)
Pease, James Roger
Issue Date
1982
Department of Study
Biochemistry
Discipline
Biochemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Chemistry, Biochemistry
Abstract
Incomplete mercuration of mRNAs followed by incomplete and controlled {('125)I}I('+) displacement of the mercury from the intermediate 5-mercuripyrimidine derivatives of the Hg-mRNAs generates {('125)I}mRNAs containing high and approximately equal specific activity ('125)I-label in both 5-{('125)I}iodouridylate and 5-{('125)I}iodocytidylate (i.e., {U,C-('125)I}mRNAs). Such {U,C-('125)I}RNAs retain their biological activity as judged by their ability to interact specifically and correctly with wheat germ 80S ribosomes in ribosome protection experiments. Further, use of such {U,C-('125)I}RNAs, along with 4 nucleotide base specific cleavage reactions, in quantitative two dimensional electrophoresis/homochromatography "fingerprint" analyses greatly facilitates one's ability to determine the RNA nucleotide sequence of the translation initiation sites of mRNAs.
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