TY - JOUR
T1 - Characterization of pURB500 from the archaeon Methanococcus maripaludis and construction of a shuttle vector
AU - Tumbula, Debra L.
AU - Bowen, Timothy L.
AU - Whitman, William B.
PY - 1997/5
Y1 - 1997/5
N2 - The complete sequence of the 8,285-bp plasmid pURB500 from Methanococcus maripaludis C5 was determined. Sequence analysis identified 18 open reading frames as well as two regions of potential iterons and complex secondary structures. The shuttle vector, pDLT44, for M, maripaludis JJ was constructed from the entire pURB500 plasmid and pMEB.2, an Escherichia coli vector containing a methanococcal puromycin-resistance marker (P. Gernhardt, O. Possot, M. Foglino, L. Sibold, and A. Klein, Mol. Gen. Genet. 221:273-279, 1900). By using polyethylene glycol transformation, M. maripaludis JJ was transformed at a frequency of 3.3 x 107 transformants per μg of pDLT44. The shuttle vector was stable in E. coli under ampicillin selection but was maintained at a lower copy number than pMEB.2. Based on the inability of various restriction fragments of pURB500 to support maintenance in M. maripaludis JJ, multiple regions of pURB500 were required. pDLT44 did not replicate in Methanococcus voltae.
AB - The complete sequence of the 8,285-bp plasmid pURB500 from Methanococcus maripaludis C5 was determined. Sequence analysis identified 18 open reading frames as well as two regions of potential iterons and complex secondary structures. The shuttle vector, pDLT44, for M, maripaludis JJ was constructed from the entire pURB500 plasmid and pMEB.2, an Escherichia coli vector containing a methanococcal puromycin-resistance marker (P. Gernhardt, O. Possot, M. Foglino, L. Sibold, and A. Klein, Mol. Gen. Genet. 221:273-279, 1900). By using polyethylene glycol transformation, M. maripaludis JJ was transformed at a frequency of 3.3 x 107 transformants per μg of pDLT44. The shuttle vector was stable in E. coli under ampicillin selection but was maintained at a lower copy number than pMEB.2. Based on the inability of various restriction fragments of pURB500 to support maintenance in M. maripaludis JJ, multiple regions of pURB500 were required. pDLT44 did not replicate in Methanococcus voltae.
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U2 - 10.1128/jb.179.9.2976-2986.1997
DO - 10.1128/jb.179.9.2976-2986.1997
M3 - Article
C2 - 9139917
AN - SCOPUS:0030903165
SN - 0021-9193
VL - 179
SP - 2976
EP - 2986
JO - Journal of bacteriology
JF - Journal of bacteriology
IS - 9
ER -