Characterization of the gene products produced in minicells by pSM1, a derivative of R100 |
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Authors: | Karen A Armstrong Hisako Ohtsubo William R Bauer Yasushi Yoshioka Chikara Miyazaki Yoshimi Maeda and Eiichi Ohtsubo |
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Institution: | (1) Institute of Applied Microbiology, University of Tokyo, Bunkyo-ku, 113 Tokyo, Japan;(2) Department of Microbiology, School of Medicine, State University of New York at Stony Brook, 11794 Stony Brook, NY, USA;(3) Present address: Department of Molecular Biology, Princeton University, 08544 Princeton, New Jersey, USA |
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Abstract: | Summary At least ten polypeptides larger than 6 kilodaltons (K) are produced in minicells from the miniplasmid pSM1 in vivo. pSM1
(5804 bp) is a small derivative of the drug resistance plasmid R100 (ca. 90 kb) and carries the R100 essential replication
region as well as some non-essential functions. Cloned restriction fragments of pSM1 and plasmids with deletions within pSM1
sequences were used to assign eight of the ten oberserved polypeptides to specific coding regions of pSM1. Two of these polypeptides
were identified as RepA1 and RepA2, proteins encoded by the essential replication region of pSM1/R100. The nucleotide sequence
consisting of 885 bp outside the essential replication region is presented here. This sequence contains an open reading frame,orf4, for a protein 22.9 K in size, and one of the pSM1-encoded polypeptides was identified as theorf4 gene product. Five additional polypeptides were shown to be the products of other open reading frames mapping outside the
essential replication region. Specific functions have been assigned to four of these polypeptides and tentatively to the fifth. |
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Keywords: | R100 pSM1 Plasmid replication proteins E coli minicells |
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