Summary: DNA fragments from the unicellular red algae Cyanidium caldarium and Porphyridium purpureum were inserted at various sites of plasmid pLG4 (pBR325 + a HindIII fragment bearing the yeast arg4 gene) with the purpose of isolating sequences supporting autonomous replication of plasmids in Saccharomyces cerevisiae. Plasmid pools were prepared in Escherichia coli then used to transform the arg4 yeast strain X3656-7 D to prototrophy. The presence of free plasmids in the yeast transformants was demonstrated by Southern blotting hybridization between yeast DNA and 32P-labelled pBR325 and by the transformation of E. coli argH with DNA from yeast transformants. Hybrid plasmids recovered from Arg+bacterial transformants transformed yeast at high frequency. They contained Aval fragments of C. caldarium total DNA, EcoRI fragments of P. purpureum satellite DNA and one BamHI fragment of P. purpureum main DNA. These new plasmids have unique restriction sites which make them convenient vectors for cloning in yeast and possibly in algae and other plants.
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