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Nucleic Acids Research, 1994, Vol. 22, No. 25 5695-5701
© 1994

Articles

The fate of deleted DNA produced during programmed genomic deletion events in Tetrahymena thermophila

Serge V. Saveliev and Michael M. Cox*

Department of Biochemistry, University of Wisconsin 420 Henry Mall, Madison, Wl 53706, USA

*To whom correspondence should be addressed

Received August 10, 1994. Revised November 4, 1994. Accepted November 4, 1994.

Thousands of DNA deletion events occur during macronuclear development In the ciliate Tetrahymena thermophila. In two deleted genomic regions, designated M and R, the eliminated sequences form circles that can be detected by PCR. However, the circles are not normal products of the reaction pathway. The circular forms occur at very low levels In conjugating cells, but are stable. Sequencing analysis showed that many of the circles (as many as 50% of those examined) reflected a precise deletion In the M and R regions. The remaining circles were either smaller or larger and contained varying lengths of sequences derived from the chromosomal DNA surrounding the eliminated region. The chromosomal junctions left behind after deletion were more precise, although deletions In either the M or R regions can generate any of several alternative junctions (1). Some new chromosomal junctions were detected in the present study. The results suggest that the deleted segment Is released as a linear DNA species that Is degraded rapidly. This species is only rarely converted to the stable circles we detect. The deletion mechanism is different from those proposed for deletion events in hypotrichous ciliates (2-4), and does not reflect a conservative site-specific recombination process such as that promoted by the bacterlophage {lambda} integrase (5).


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