Characterization of a chemically synthesized RNA having the sequence of the yeast initiator tRNAMet

FEBS Letters

Keith, Gérard²  Perreault, Jean-Pierre³  Bratty, John³  Ogilvie, Kelvin K.¹  Wu, Taifeng¹  Cedergren, Robert³  Nicoghosian, Krikor³ 

[1] Department of Chemistry, McGill University, Montréal, Qué, H3A 2K6, Canada;Institut de Biologie Moléculaire et Cellulaire du CNRS, F-67084 Strasbourg, France;Département de Biochimie, Université de Montréal, Montréal, Qué, H3C 3J7, Canada

关键词: RNA;    Chemical synthesis;    Purification;    Aminoacylation;    PAGE;    polyacrylamide gel electrophoresis;    BND-cellulose;    benzoylated naphthoylated diethylaminoethylcellulose;    EDTA;    ethylenediaminetetraacetate;    pCp;    3';    5'-cytidine diphosphate;    Phy M and B.c.;    RNases from Physarum and Bacillus cereus;    respectively;    TBAF;    tetrabutylammonium fluoride. One OD260 is the quantity of material that;    when dissolved in 1 ml;    gives an A260 of one with a 1 cm pathlength;   

DOI  :  10.1016/0014-5793(90)81118-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A 75-unit long oligoribonucleotide corresponding to the sequence of the Saccharomyces cerevisiae initiator tRNA was synthesized chemically. The crude RNA was purified, and the sequence was verified by RNA sequencing techniques. A particularly useful purification step involved hydrophobic chromatography on BND-cellulose. The purified RNA could be aminoacylated to 28% of a bona fide initiator tRNA^Metsample and threonylated to 76 % of the level observed with native tRNA^fMet from E. coli.

【 授权许可】

Unknown