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A purification techniques was developed to obtain a preparation of the bacteriophage T4 RNA-ligase (EC 6.5.1.3) free of contaminating nuclease activities. It includes fractioning on phosphocellulose and DEAE-cellulose, chromatography on DEAE-cellulose, isoelectric sedimentation, gel filtration through Sephadex G-100 and chromatography on hydroxylapatite and aminohexyl-sepharose. The enzyme yield amounts to 30-37%. By means of biochemical tests, the preparation of RNA-ligase was found to be good for molecular biology and gene engineering, in particular for such areas where oligodeoxyribonucleotides and high molecular-weight RNA are used as substrates. The ligating ability of the enzyme was demonstrated by means of the preparative synthesis of nonaribonucleotide ApUpG(pU)6 from trinucleoside diphosphate ApUpG and hexoribouridylic acid, as well as by insertion of [5'-32P]-labelled cytidine-3',5'-diphosphate in the 3'-end of the bacteriophage MS 2 RNA.
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
October 1972,
Proceedings of the National Academy of Sciences of the United States of America,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
May 1976,
Archives of biochemistry and biophysics,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
January 1984,
Prikladnaia biokhimiia i mikrobiologiia,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
January 1987,
Molekuliarnaia biologiia,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
January 1984,
Molekuliarnaia biologiia,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
February 2004,
Virology,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
September 1977,
Nucleic acids research,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
December 2002,
BioTechniques,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
January 1976,
Biochemical and biophysical research communications,
M M Baklanov, and
I A Riazankin, and
A S Butorin, and
Iu S Nechaev, and
V I Iamkovoĭ
December 2003,
Nucleic acids research,
