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A method for the analysis of tRNA patterns in Drosophila melanogaster using an amino acid analyser

✍ Scribed by R.A. Davey; A.J. Howells

Publisher: Elsevier Science
Year: 1974
Tongue: English
Weight: 578 KB
Volume: 60
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(74)90257-7

No coin nor oath required. For personal study only.

✦ Synopsis

Using tRNA and aminoacyl-tRNA synthetase preparations from Drosophila melanogaster, a method has been developed for simultaneously estimating levels of at least 15 different species of aminoacyl-tRNA.

"C-labeled aminoacyl-tRNA, which is formed during a single incubation of tRNA with a mixture of 15 'YXabeled amino acids, is purified, hydrolysed, and the composition of the mixture of "C-labeled amino acids so obtained is determined using an Amino Acid Analyser.

The sensitivity of the method and the reproducibility of the results obtained are such that it is suitable for detecting changes in tRNA patterns in comparative studies.

Methods

Reagents. 8-Hydroxyquinoline was obtained from British Drug Houses Ltd.; disodium naphthalene-1, 5-disulphonate from Fluka; sodium toluene-p-sulphonate from Eastman Kodak Co. ; 14C-labeled amino acids from the Radiochemical Centre, Amersham, and from C.E.A. (French Atomic Energy Commission) ; phenol and m-cresol from May and Baker 469

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