Characterization of Rhodococcus-E. coli shuttle vector pNC9501 constructed from the cryptic plasmid of a propene-degrading bacterium.
Rhodococcus-E. coli shuttle vector pNC9501 was constructed using circular cryptic plasmid pNC903 from propene-degrading Rhodococcus ruber P-II-123-1. Sequence analysis of pNC903 revealed two open-reading frames encoding the replication proteins Reps A and B. In the amino acid sequence of the putative Rep B, a helix-turn-helix motif, which is responsible for the binding of DNA, was found. Sequencing of the upstream region of the putative Rep A and incompatibility tests revealed that pNC903 is a Mycobacterium-derived pAL5000-related plasmid. pNC9501 could also be transformed into Mycobacterium sp. showing good segregation stability (<0.1% plasmid loss/generation) in the absence of selective pressure.
Matsui
Toru
T
Center of Molecular Biosciences, University of the Ryukyus, 1 Sembaru, Okinawa 903-0213, Japan. tmatsui@comb.u-ryukyu.ac.jp
Saeki
Hisashi
H
Shinzato
Naoya
N
Matsuda
Hitoshi
H
D000475:Alkenes / Q000378:metabolism; D000595:Amino Acid Sequence; D004926:Escherichia coli; D005822:Genetic Vectors / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D010957:Plasmids / Q000096:biosynthesis / Q000235:genetics; D012240:Rhodococcus / Q000235:genetics*; D014169:Transformation, Bacterial / Q000235:genetics
D000475:Alkenes; C013658:propylene