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Nucleotide Sequence Analysis of Cryptic Plasmid pAM5 from Acidiphilium Multivorum

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Title Nucleotide Sequence Analysis of Cryptic Plasmid pAM5 from
Acidiphilium Multivorum
 
Creator Singh, Samarendra K
Banerjee, Pataki C
 
Subject Drug Development/Diagnostics & Biotechnology
 
Description Plasmid pAM5 of Acidiphilium multivorum JCM-8867 has been completely sequenced by initial cloning of HindIII–PstI
fragments followed by primer walking. It has a size of 5161 bp and single site for several restriction enzymes as revealed by
DNA sequencing. Sequence analysis predicts five putative open reading frames. ORF1 and ORF3 show significant identity
with various plasmid encoded mobilization (Mob) and replication initiation (Rep) proteins, respectively. The putative Mob
protein has several characteristics of the MOBQ family having the motifs with conserved amino acid residues. Upstream of
the Mob ORF, there exists a 34 bp oriT region having a nic consensus sequence. The constructed plasmid pSK1 bearing
pAM5 mob region can be mobilized to Escherichia coli in presence of conjugative plasmid pRK2013. The replication module
comprises of several DnaA like boxes, several perfect direct and inverted repeats, a potential prokaryotic promoter and
putative rep gene. The rep module is very similar to several theta replicating iteron family plasmids, suggesting pAM5 replication
to follow the same course. Any phenotypic character determinant (e.g., metal resistance, antibiotic resistance etc.)
gene is absent in pAM5, suggesting this plasmid to be cryptic in nature. However, a pAM5 derivative plasmid named
pSK2, containing the putative pAM5 rep region, can replicate and be stably maintained in Acidiphilium, Acidocella,
and E. coli strains; it can also carry foreign DNA fragments. Thus, pSK2 could serve as a cloning shuttle vector between
these bacteria. It was observed that pAM5 Rep is essential for pSK2 to replicate in acidophiles. In its natural host,
A. multivorum JCM-8867, pAM5 maintains a copy number of 50–60, and its derivative pSK2 maintains a comparatively,
higher copy number in E. coli than in acidophiles.
 
Publisher Elsevier
 
Date 2007
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/1152/1/PLASMID_58(_2)_101%2D114;2007[38].pdf
Singh, Samarendra K and Banerjee, Pataki C (2007) Nucleotide Sequence Analysis of Cryptic Plasmid pAM5 from Acidiphilium Multivorum. Plasmid, 58 (2). pp. 101-114.
 
Relation http://dx.doi.org/10.1016/j.plasmid.2007.01.005
http://www.eprints.iicb.res.in/1152/