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Nucleic Acids Research, 1983, Vol. 11, No. 7 2053-2064
© 1983

MOLECULAR BIOLOGY

Using iodinated single–stranded M13 probes to facilltate rapid DNA sequence analysis – nucleotide sequence of a mouse lysine tRNA gene

Jang H. Han and John D. Harding*

922 Fairchild Center, Department of Biological Sciences, Columbia University New York, NY 10027, USA

*To whom to address correspondence

Received December 29, 1983. Revised March 7, 1983. Accepted March 7, 1983.

From a recombinant lambda phage, we have determined a 387 bp sequence containing a mouse lysine tRNA gene. The putative lys tRNA (anticodon UUU) differs from rabbit liver lys tRNA at five positions. The flanking regions of the mouse gene are not generally homologous to published human and Drosophila lys tRNA genes. However, the mouse gene contains a 14 bp region comprising 13 A–T base pairs, 30–44 bp from the 5' end of the coding region. Cognate A–T rich regions are present in human and Drosophila genes. The coding region is flanked by two 11 bp direct repeats, similar to those associated with alu family sequences.

The sequence was determined by a "walking" protocol that employs, as a novel feature, iodinated single–stranded M13 probes to identify M13 subclones which contain sequences partially overlapping and contiguous to an initially determined sequence. The probes can also be used to screen lambda phage and in Southern and dot blot experiments


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