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AssigneeArizona Board of Regents on behalf of Arizona State University
InventorsBo Wang, Weiwen Zhang, Deirdre Meldrum
Estimating the term…
Methods of pre-methylation of foreign DNA to improve genetic transformation in cyanobacterium. Two Type II methyltransferase-encoding genes, i.e., M (sll0729) and C (slr0214), were cloned from the chromosome of Synechocystis sp. PCC 6803 (hereafter Synechocystis 6803) and expressed in E. coli that harbors the integrative plasmid pBS-SPtK or pJU105. After pre-methylation in E. coli, the integrative plasmids were extracted and used for transformation of Synechocystis 6803. The results showed that expression of slr0214 in the integrative-plasmid-harboring E. coli cells before DNA preparation resulted in orders of magnitude higher efficiency in the following integrative transformation of Synechocystis 6803.
A method utilizing pre-methylated DNA that has been methylated in situ and then isolated from a recombinant host cell for enhanced efficiency transformation of cyanobacteria versus transformation with DNA that has not been pre-methylated and isolated from said recombinant host cell, comprising the step of:
A method utilizing pre-methylated DNA that has been methylated in situ and then isolated from a recombinant host cell for enhanced efficiency transformation of a cyanobacteria cell versus transformation with DNA that has not been pre-methylated and isolated from said recombinant host cell, comprising transforming said cyanobacteria cell by adding DNA comprising a plasmid mixture to a culture that contains said cyanobacteria cell under conditions and for a time sufficient to effect transformation, wherein said DNA comprising said plasmid mixture has been methylated in, and isolated from, said recombinant host cell, and wherein said DNA integrates into a genome of the cyanobacteria cell.
Dependencies are inferred from the claim text.