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Nucleic Acids Research, 1988, Vol. 16, No. 9 4137-4153
© 1988

Articles

Heterologous 5' flanking regions do not support in vitro template activity of a Drosophila melanogaster tRNAVal3b gene

Ron M. Mackay+, Daniel Horvath, Loverne Duncan and George B. Spiegelman*

Departments of Microbiology and Medical Genetics, University of British Columbia Vancouver, British Columbia, V6T 1W5, Canada

*To whom correspondence should be addressed

Received June 19, 1987. Revised February 24, 1988. Accepted February 24, 1988.

The 5' and 3' structure of a Drosophila tRNAVal3b gene was investigated to examine the defect which caused the extremely low in vitro transcription template activity of the gene. Recombinant genes were constructed linking 5' and 3' flanking regions from tRNA genes which were active in vitro templates (tRNAVal4, tRNAArg, tRNASer7) to the tRNAVal3b gene. None of the recombinant genes were effective in vitro templates. The defect in tRNAVal3b was demonstrated to reside in the 5' flanking region of the gene and deletion analysis indicated that no specific transcription inhibitor sequence was present 5' to the gene. The data suggest that the effect of 5' flanking sequences on in vitro transcription of the tRNAVal3b gene requires a specific relationship between the tRNA gene and the flanking sequence.

+Present address: National Research Council of Canada, Room 3142, 100 Sussex Dr., Ottawa, Ontario, K1A OR6, Canada


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