TY - JOUR
T1 - A genetic system for Archaea of the genus Methanosarcina
T2 - Liposome-mediated transformation and construction of shuttle vectors
AU - Metcalf, William W.
AU - Zhang, Jun Kai
AU - Apolinario, Ethel
AU - Sowers, Kevin R.
AU - Wolfe, Ralph S.
PY - 1997/3/18
Y1 - 1997/3/18
N2 - New methods that allow, for the first time, genetic analysis in Archaea of the genus Methanosarcina are presented. First, several autonomously replicating plasmid shuttle vectors have been constructed based on the naturally occurring plasmid pC2A from Methanosarcina acetivorans. These vectors replicate in 9 of 11 Methanosarcina strains tested and in Escherichia coli. Second, a highly efficient transformation system based upon introduction of DNA by liposomes has been developed. This method allows transformation frequencies of as high as 2 x 108 transformants per microgram of DNA per 109 cells or ≈20% of the recipient population. During the course of this work, the complete 5467-bp DNA sequence of pC2A was determined. The implications of these findings for the future of methanoarchaeal research are also discussed.
AB - New methods that allow, for the first time, genetic analysis in Archaea of the genus Methanosarcina are presented. First, several autonomously replicating plasmid shuttle vectors have been constructed based on the naturally occurring plasmid pC2A from Methanosarcina acetivorans. These vectors replicate in 9 of 11 Methanosarcina strains tested and in Escherichia coli. Second, a highly efficient transformation system based upon introduction of DNA by liposomes has been developed. This method allows transformation frequencies of as high as 2 x 108 transformants per microgram of DNA per 109 cells or ≈20% of the recipient population. During the course of this work, the complete 5467-bp DNA sequence of pC2A was determined. The implications of these findings for the future of methanoarchaeal research are also discussed.
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U2 - 10.1073/pnas.94.6.2626
DO - 10.1073/pnas.94.6.2626
M3 - Article
C2 - 9122246
AN - SCOPUS:0030900723
SN - 0027-8424
VL - 94
SP - 2626
EP - 2631
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 6
ER -