-transparent.png){kind=logo}
We have undertaken a search for autonomously replicating (ARSs) from Kluyveromyces lactis chromosomal DNA able to sustain plasmid replication in K. lactis and in Saccharomyces cerevisiae. The discovery of such sequences might be interesting for the comparison of ARSs from different sources and possibly useful for the construction of multivalent vectors. HindIII fragments from K. lactis chromosomal DNA were inserted in the YIp5 plasmid (lacking an origin of replication) and the resulting chimaeric plasmids were selected for the ability to transform S. cerevisiae. Four plasmids were identified and further analysed. Two contained the same 1.8 kb K. lactis fragment and transformed both K. lactis and S. cerevisiae with the same efficiency and stability, whereas the third transformed only S. cerevisiae and the fourth transformed K. lactis with a higher efficiency than S. cerevisiae. A detailed study was performed on the 1.8 kb fragment which exhibited ARS function in both yeasts. The fragment was subcloned using different restriction enzymes and Bal31 exonuclease. Subclones were tested for ARS function. ARS activities in the two yeasts were localized in the same 100 bp region. Sequencing demonstrated the presence in this region of the dodecanucleotide 5'ATTTATTGTTTT3' differing from the ARS core consensus of S. cerevisiae only by a T insertion. A similar nucleotide sequence is present in the putative replication origin of the 2 mu-like plasmid pKD1 which stably replicates in K. lactis. Homologies with ARSs from S. cerevisiae were also found in the regions flanking the above-mentioned dodecanucleotide.
L Fabiani, and
M Aragona, and
L Frontali
January 1984,
Nucleic acids research,
L Fabiani, and
M Aragona, and
L Frontali
April 1990,
Molecular & general genetics : MGG,
L Fabiani, and
M Aragona, and
L Frontali
March 1988,
Plant molecular biology,
L Fabiani, and
M Aragona, and
L Frontali
October 1989,
Genomics,
L Fabiani, and
M Aragona, and
L Frontali
October 1990,
Molecular and cellular biology,
L Fabiani, and
M Aragona, and
L Frontali
March 1989,
Molecular and cellular biology,
L Fabiani, and
M Aragona, and
L Frontali
December 1995,
Genome,
L Fabiani, and
M Aragona, and
L Frontali
February 1990,
Molecular & general genetics : MGG,
L Fabiani, and
M Aragona, and
L Frontali
February 1988,
Proceedings of the National Academy of Sciences of the United States of America,
L Fabiani, and
M Aragona, and
L Frontali
June 1993,
Proceedings of the National Academy of Sciences of the United States of America,
