Transformation methods for halophilic archaebacteria.
We present a practical description of polyethylene glycol mediated spheroplast transformation of Halobacterium halobium and Halobacterium volcanii. This method has been applied to phage DNA transfection, plasmid DNA transformation, and transformation with linear fragments of high molecular weight genomic DNA. Efficient spheroplast regeneration allows uncomplicated recovery of transformed progeny. Transformations can be performed equally well using fresh or frozen cell preparations. These methods should find application in molecular cloning, genetic fine mapping, and strain construction.
Cline
S W
SW
Department of Biochemistry, Dalhousie University, Halifax, N.S., Canada.
Lam
W L
WL
Charlebois
R L
RL
Schalkwyk
L C
LC
Doolittle
W F
WF
D001105:Archaea / Q000235:genetics; D001435:Bacteriophages / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005821:Genetic Techniques; D006217:Halobacterium / Q000235:genetics*; D010957:Plasmids; D011092:Polyethylene Glycols; D013104:Spheroplasts; D014162:Transfection; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D004279:DNA, Viral; D011092:Polyethylene Glycols