Abstract
Evidence is presented for the autonomous replication of recombinant plasmids in Mucor circinelloides Leu+ transformants. Plasmids consisting of a unique fragment of Mucor DNA inserted into YRp17 or pBR322 (i) give a high frequency of transformation (up to 7800 Leu+ transformants per μg DNA), (ii) are mitotically unstable, (iii) can be reisolated in an unmodified form from uncut transformant DNA, and (iv) are present as discrete extrachromosomal DNA molecules (as detected by Southern hybridisation). No integration of the transforming DNA can be detected in Mucor transformants containing free plasmid molecules. This excludes the possibility that free plasmids are simply excision products resulting from a reversal of the integration process. Sub-cloning of the recombinant plasmids and analysis of subsequent Leu+ transformants show that autonomous replication is independent of the vector sequences and locates a Mucorars within a 4.4 kb PstI fragment of the insert DNA.
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