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The primary structure of leucine transfer rna from cow mammary gland oligo nucleotides of t 1 rnase hydrolysate the reconstruction of the total nucleotide sequence


, : The primary structure of leucine transfer rna from cow mammary gland oligo nucleotides of t 1 rnase hydrolysate the reconstruction of the total nucleotide sequence. Bioorganicheskaya Khimiya 10(1): 58-67

The oligonucleotides obtained by digestion of leucine transfer RNA from cow mammary gland with T1 RNase were separated by micro-column chromatography on DEAE-cellulose in 7 M urea at pH 7.5 and 3.7, and in addition on Dowex 1 .times. 2. The digest consisted of 18 individual components, the larger being a tridecanucleotide. Micro-column chromatography of nucleotides on anion-exchanger AG 1 .times. 8 and nucleosides on Aminex A-6 was used to determine the base composition of the oligonucleotides. The oligonucleotide structure was established using terminal analysis, hydrolysis by pyrimidyl and U2-RNases and incomplete hydrolysis by snake venom phosphodiesterases. The total primary structure of leucine transfer RNA was derived from overlapping fragments isolated after its complete hydrolysis with pancreatic and T1 RNase and using data obtained on S1-nuclease digestion of tRNA. The methods of rapid gel-sequencing were also employed for checking the nucleotide sequence of leucine transfer RNA from cow mammary gland.

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