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Title: Studies on the structure and function of E. coli phenylalanine transfer RNA
Author: Lowdon, Margaret
ISNI:       0000 0001 3613 2468
Awarding Body: University of Glasgow
Current Institution: University of Glasgow
Date of Award: 1976
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Two different methods have been used for the purification of E. coli TRNA[Phe2]. Both methods involved initial fractionation of unpurified E. coli tRNA (either unlabelled, or uniformly labelled with [32]P) on Benzoylated DEAE-cellulose. The fractions containing TRNA[Phe2] (the second phenylalanine isoacceptor tRNA eluted from the Benzoylated DEAE-cellulose column on increasing the NaC1 concentration of the eluate) were then fractionated either after charging of the tRNA with phenylalanine on another Benzoylated DEAE-cellulose column, or Rhe on an RPC-5 column. After deacylation of purified Phe-tRNA[Phe2] the tRNA[Phe2] could not be charged with phenylalanine to the original extent. Purified tRNA[Phe2] obtained from an RPC-5 column was found to lose phenylalanine accepting activity on storage. Attempts were made to renature both inactivated forms of tRNA[Phe2] but they were not successful. Attempts to fully charge the inactivated tRNA[Phe2] by increasing the ligase concentration in the assay or by preincubating the tRNA[Phe2] with the ligase were equally unsuccessful. However, after investigation of the stability of tRNA[Phe2] under various conditions, it was found to be most stable in 10mr1 MgC12, 10mM tris-HC1, pH 7.0, in the presence of an equal amount of unpurified E. coli tRNA. The purified E. coli tRNA[Phe2] isolated was found to give identical T1 and Pancreatic RNase fingerprints to the E. coli tRNA[Phe] whose sequence was elucidated by Barrel & Sanger (1969).
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available