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Nucleic Acids Research, 1983, Vol. 11, No. 13 4379-4389
© 1983

MOLECULAR BIOLOGY

Molecular cloning and sequencing of pheU, a gene for Escherichia coli tRNAPhe

Ira Schwartz1, Robin-Ann Klotsky1, Dirk Elseviers2, Patricia J. Gallagher2, Manuel Krauskopf3,*, M.A.Q. Siddiqui3, James F.H. Wong4 and Bruce A. Roe4

1Departments of Biochemistry, New York Medical College Valhalla, NY 10595 2Microbiology, New York Medical College Valhalla, NY 10595 3Roche Institute of Molecular Biology Nutley, NJ 07110 4Department of Chemistry, University of Oklahoma Norman, OK 73019, USA

Received April 6, 1983. Revised June 1, 1983. Accepted June 1, 1983.

A recombinant plasmid (designated pID2) carrying the E. coli gene for tRNAPhe has been isolated from a plasmid bank constructed by the ligation of a total EcoRI digest of E. coli K12 DNA into the EcoRI site of pACYC184 DNA. The plasmid was selected by virtue of its ability to complement a temperature-sensitive lesion in the gene (PheS) for the {alpha}-subunit of phenylalanyl-tRNA synthetase. Crude tRNA isolated from such transformants exhibited elevated levels of phenylalanine acceptor activity. The tRNAPhe gene has been localized within the first 300 base pairs of a 3.6 kb SalI fragment of pID2. The sequence of the gene and its flanking regions is presented.

*Permanent Address: inst. de Bioquimica, Fac. de Ciencias, Univ. Austral de Chile, Vaidivia, Chile.


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