Patent · US Active

Premethylation of DNA for high efficiency transformation of cyanobacteria

US10378019B2 · kind B2 · utility

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20Claims
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Key dates

Filing dateSep 1, 2015
Grant dateAug 13, 2019
Priority date
Expiry dateSep 1, 2035

Classification

  • Technology area (CPC C)Chemistry; Metallurgy
  • CPC primaryC12N15/74
  • WIPO fieldBiotechnology
  • WIPO sectorChemistry

Abstract

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.

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