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Stability of cloned Brassica napus chloroplast DNA fragments in the cyanobacterium Anacystis nidulans R2
Authors:Wei Xiao
Institution:(1) Department of Biology, University of Saskatchewan, S7N OWO Saskatoon, Saskatchewan, Canada
Abstract:Summary Brassica napus (cv. ldquoTritonrdquo) chloroplast (cp) DNA BamHI gragments were inserted into a bacteria-cyanobacteria shuttle vector pCB4. The chloroplast genomic library was screened in Escherichia coli and 28 individual clones, which represent 94% of the total chloroplast genome, were isolated. Cyanobacterium Anacystis nidulans R2 was transformed with each member of the clone bank by selection for ampicillin resistance. A study of transformation efficiency showed dramatic variation (up to 200-fold) among recombinant clones. Furthermore, plasmid DNA reisolated from some cyanobacterial transformants exhibited instability. Variations in transformation efficiency and plasmid instability were shown to be DNA sequence specific. B. napus cpDNA clones were thus classified into three types according to their stability in the cyanobacterial host.
Keywords:Brassica napus  Chloroplast genome  Anacystis nidulans  Cyanobacterial transformation  Plasmid stability
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