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Nucleic Acids Research, 1984, Vol. 12, No. 15 6169-6178
© 1984

MOLECULAR BIOLOGY

Characterization of deletion derivatives of an autonomously replicating Neurospora plasmid

Lori L. Stohl, Robert A. Akins and Alan M. Lambowitz*

E.A.Doisy Department of Biochemistry, Saint Louis University School of Medicine St. Louis MO 63104, USA

*To whom correspondence should be addressed

Received March 26, 1984. Revised June 29, 1984. Accepted June 29, 1984.

We previously described two plasmids that replicate autonomously in both Neurospora and E. coli (Stohl arid Lambowitz, Proc. Natl. Acad. Sci., U.S.A 80, 1058–1062, 1983). One plasmid, pALS1, consists of the Neurospora qa-2+ gene (3 kb Hind III-fragment), the mitochondrial plasmid from N. intermedia strain P405-Labelle, and E. coli plasmid pBR325. The other, pALS2, is a putative deletion derivative of pALS1 that lacks most or all of the Labelle insert and that was repeatedly recovered from Neurospora transformants. We have now sequenced the region encompassing the deletion in five pALS2 plasmids isolated independently in two different laboratories. All five plasmids are identical in this region and completely lack the Labelle insert. We have also characterized an additional deletion derivative that retains a small (approximately 0.5 kb) segment of the Labelle insert. The results for pALS2 suggest that pBR325 plus the ga-2+ segment constitute a Neurospora replicon.


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