Methanogenic - University of Maryland
Methanogenic - University of Maryland
Methanogenic - University of Maryland
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Hybrid Shuttle Vector<br />
trpA terminator<br />
oriRK6<br />
mcr terminator<br />
pac<br />
mcr promoter<br />
EcoRV (183) BamHI (187) XhoI (265)<br />
bla<br />
lacZ<br />
PWM315<br />
8745bp<br />
pC2A<br />
Ecl136II (233)<br />
EcoICRI (233)<br />
SstI (235)<br />
SacI (235)<br />
Asp718I (240)<br />
Acc65I (240)<br />
KpnI (244)<br />
SphI (253)<br />
ClaI (256)<br />
Metcalf et al., 1997<br />
Shuttle Vector Hybrid Constructs<br />
• pJK series Integration vector Methanosarcina spp.<br />
• pWM series Autonomous vector Methanosarcina spp.<br />
• pWAY1/2 Integration vector Methanosarcina<br />
mazei<br />
• pMIP1 Integration vector Methanococcus voltae<br />
• pKAS Integration vector Methanococcus<br />
maripaludis<br />
• pDLT44 Autonomous vector Methanococcus<br />
maripaludis<br />
Transformation Protocols<br />
• Natural transformation (M. voltae) < 7 x 10 2 /µg<br />
• Electroporation (M. voltae) < 7 x 10 2 /µg<br />
• PEG (M. maripaludis) < 2 x 10 7 /µg<br />
• Liposome (Methanosarcina spp.) < 2 x 10 9 • Natural transformation (M. voltae) < 7 x 10<br />
/µg<br />
2 /µg<br />
• Electroporation (M. voltae) < 7 x 10 2 /µg<br />
• PEG (M. maripaludis) < 2 x 10 7 /µg<br />
• Liposome (Methanosarcina spp.) < 2 x 10 9 /µg
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