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KMID : 0578319930030040373
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Molecules and Cells
1993 Volume.3 No. 4 p.373 ~ p.378
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Replication of the Lactococcal Plasmid pGKV2.1 in E. coli
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Jeong, Jin Yong
Seo, Hak Soo/Kim, Ho Yeon/Choi, Choong Kuk/Cho, Moo Je/Bahk, Jeong Dong
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Abstract
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The broad-host-range plasmid pGKV21 of Lactococcus lactic has been shown to replicate in both gram-positive and gram-negative bacteria. In E. soli, pGKV21 replicated via a single stranded circular DNA intermediate as other plasmids isolated from gram-positive bacteria, suggesting that this plasmid uses the rolling circle mode of replication. In vitro replication of plasmid pGKV21 was examined using extracts prepared from plasmid-free and -containing E. coli cells. It was found that the specific plasmid-encoded factor was needed for the replication of the lactococcal plasmid pGKV21. The putative minus origin of pGKV21 was determined by their capability for the initiation of complementary strand synthesis of a mutant M13 phage vector whose minus origin had been largely deleted. The growth activity of a recombinant M13 phage carrying the 228 by region between DdeI-Dral of pGKV21 could partially recover the defect in replication of the mutant M13 phage. This region contained a large axis of dyad symmetry and showed orientation-dependent activity in replication. The putative minus origin detected in pGKV21 did share some sequence homologies with the complementary strand origin of the E coli single stranded DNA bacteriophage (DX174 but not with those of other plasmids from gram-positive bacteria.
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