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Molecular Characterization of Plasmid from Bifidobacterium longum  

Park, Myeong-Soo (Research Center, BIFIDO Co., Ltd.)
Moon, Hye-Won (Department of Food Science and Technology, Seoul National University)
Ji, Geun-Eog (Research Center, BIFIDO Co., Ltd., Department of Food and Nutrition, Seoul National University)
Publication Information
Journal of Microbiology and Biotechnology / v.13, no.3, 2003 , pp. 457-462 More about this Journal
Abstract
The complete nucleotide sequence of a plasmid, pMG1, isolated from Bifidobacterium longum MG1 has been determined. This plasmid, composed of 3,862 base pairs with 65.1% of G+C content. harbors two major open reading frames (ORF) encoding putative proteins of 29 kDa (ORF I) and 71 kDa (ORF II). ORF I showed relatively high amino acid sequence homology with replication proteins of other plasmids from Gr Im-positive and -negative bacteria. Upstream of ORF I, four sets of tandem repeat sequences resembling the iteron structure of related plasmids were found. S1 endonuclease treatment and Southern blot analysis revealed that pMG1 accumulates single-stranded DNA (ssDNA) intermediate, which indicate i the rolling circle replication (RCR) mechanism of this plasmid. Homology search indicated that ORF II encodes plasmid mobilization protein, and the presence of highly conserved oriT sequence in the upstream of this gene supported this assumption. RT-PCR showed that only ORF I is expressed in vivo. Based on these results, pMG 1 was exploited to construct a shuttle vector, pBES2. It was successfully transformed into Bifidobacterium and maintained stably.
Keywords
Bifidobacterium longum MG1; plasmid; sequence analysis; shuttle vector; electroporation;
Citations & Related Records
Times Cited By KSCI : 3  (Citation Analysis)
Times Cited By Web Of Science : 12  (Related Records In Web of Science)
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