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Characterization of the minimal origin required for replication of the streptococcal plasmid plP501 in Bacillus subtilis
Author(s) -
Brantl S.,
Behnke D.
Publication year - 1992
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1992.tb01785.x
Subject(s) - biology , plasmid , autonomously replicating sequence , origin of replication , genetics , bacillus subtilis , gene , transformation (genetics) , dna , microbiology and biotechnology , nucleic acid sequence , dna replication , bacteria
Summary By using deletional analysis the origin of replication, oriR , of the streptococcal plasmid plP501 in Bacillus subtilis has been mapped at a position immediately downstream of the repR gene. Determination of both the right and left border of oriR allowed the definition of a sequence of a maximum of 52 nucleotides which theoretically constitutes the minimal origin of replication. Recently, the start point of leading‐strand synthesis of the closely related plasmid pAMβ1 has been mapped at a position which is located exactly in the middle of this sequence (Bruand et al. , 1991). The function of ohR did not depend on its location downstream of the repR gene. Translocation of oriR containing fragments to other regions of the plasmid proved to be possible. The smallest translocated fragment that still reconstituted autonomous replication was 72bp in size. This fragment was also active in directing the replication of an Escherichia coli plasmid in B. subtilis when the RepR protein was supplied in trans from a repR gene integrated into the host chromosome. The transformation efficiency of plasmids carrying translocated oriR fragments showed a certain dependence on the fragment length and orientation. The DNA sequence of oriR included an inverted repeat, both branches of which appeared to be essential for oriR function. The repeats of oriR shared sequence similarity with a repeat located upstream of promoter pll, which has been suggested to be involved in autoregulation of repR expression.