TY - JOUR
T1 - Physical relationship between a gene and its origin of replication in Physarum polycephalum
AU - Benard, Marianne
AU - Pierron, Gérard
N1 - Funding Information:
We gratefully acknowledge the technical assistance of Jacqueline Pedron. We thank G. Lemieux and D. Pallotta (Universite Laval, Quebec) for generously providing many cDNA clones and E. Puvion, director of our laboratory, for his constant support. M. B. is a fellow of the Institut de Formation Superieure Biomedicale (Villejuif). 6. P. is supported by Grant 1301 from the Association pour la Recherche sur le Cancer.
PY - 1990/1/1
Y1 - 1990/1/1
N2 - Taking advantage of the natural synchrony of the S-phase within the plasmodium of Physarum polycephalum, we extracted highly synchronous DNA samples at precise time points in early S-phase. We then separated, by electrophoresis under denaturating conditions, the newly synthesized DNA strands of the nascent chromosomal replicons from the parental DNA template. Using the cDNA clone of the early-replicating LAV1-2 gene as a probe, we could establish by filter hybridization that the elongation rate of the replicon which encompasses this gene is constant, at a rate of 1 kb/min during the first 30 min of S-phase. The smallest replication intermediate (RI) that we have detected by probing with the LAV1-2 cDNA was 5 kb long, suggesting that the LAV1-2 gene and its origin of replication are closely associated within the chromosome. This procedure should facilitate the mapping of replication origins within the genome of Physarum.
AB - Taking advantage of the natural synchrony of the S-phase within the plasmodium of Physarum polycephalum, we extracted highly synchronous DNA samples at precise time points in early S-phase. We then separated, by electrophoresis under denaturating conditions, the newly synthesized DNA strands of the nascent chromosomal replicons from the parental DNA template. Using the cDNA clone of the early-replicating LAV1-2 gene as a probe, we could establish by filter hybridization that the elongation rate of the replicon which encompasses this gene is constant, at a rate of 1 kb/min during the first 30 min of S-phase. The smallest replication intermediate (RI) that we have detected by probing with the LAV1-2 cDNA was 5 kb long, suggesting that the LAV1-2 gene and its origin of replication are closely associated within the chromosome. This procedure should facilitate the mapping of replication origins within the genome of Physarum.
UR - http://www.scopus.com/inward/record.url?scp=0025125714&partnerID=8YFLogxK
U2 - 10.1016/0014-4827(90)90309-X
DO - 10.1016/0014-4827(90)90309-X
M3 - Article
C2 - 1967581
AN - SCOPUS:0025125714
SN - 0014-4827
VL - 186
SP - 299
EP - 305
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -