Details

Autor(en) / Beteiligte

• Stohl, Lori L.
• Lambowitz, Alan M.

Titel

Construction of a Shuttle Vector for the Filamentous Fungus Neurospora crassa

Ist Teil von

• Proceedings of the National Academy of Sciences - PNAS, 1983-02, Vol.80 (4), p.1058-1062

Ort / Verlag

United States: National Academy of Sciences of the United States of America

Erscheinungsjahr

1983

Quelle

MEDLINE

Beschreibungen/Notizen

• We have constructed a recombinant plasmid, pALS-1, that replicates autonomously in both Neurospora and Escherichia coli. pALS-1 consists of the mitochondrial plasmid from Neurospora strain P405-Labelle, the Neurospora qa-2+ gene, and E. coli plasmid pBR325. pALS-1 transforms the Neurospora qa-2+ gene at frequencies 5- to 10-fold higher than those for plasmids that transform mainly by integration. When E. coli was transformed with DNA from Neurospora transformants, we recovered not only pALS-1 but also a smaller plasmid, pALS-2, which had undergone deletion of most and possibly all Labelle sequences, and the immediately flanking sequences in pBR325. pALS-2 also appears to replicate autonomously in Neurospora, but less efficiently than does pALS-1. Southern blots show that free pALS-1 and pALS-2 are present in nuclear and cytosolic (supernatant from high-speed centrifugation) fractions of Neurospora transformants and that small, variable proportions of the plasmids also can be detected in mitochondria. pALS-1 and pALS-2 constitute putative shuttle vectors for Neurospora.