Efficient delivery of large DNA from Escherichia coli to Synechococcus elongatus PCC7942 by broad-host-range conjugal plasmid pUB307

Mitsuhiro Itaya, Hiroko Kusakabe, Mitsuru Sato, Masaru Tomita, Rintaro Sato

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Synechococcus elongatus PCC7942, a cyanobacterium that uses light and carbon dioxide to grow, has a high ability to incorporate DNA by transformation. To assess the effective delivery of large DNA in plasmid form, we cloned the endogenous plasmid pANL (46.4 kbp) into a BAC vector of Escherichia coli. The plasmid p38ANL (54.3 kbp) replaced the native plasmid. To assess the delivery of larger DNA into PCC7942, p38ANL was fused to the broad-host-range conjugal transfer plasmid pUB307IP (53.5 kbp). The resulting plasmid pUB307IP501 (107.9 kbp) was transmitted from E. coli to PCC7942 by simple mixing of donor and recipient cultures. PCC7942 transcipients possessed only pUB307IP501, replacing the preexisting pANL. In contrast, the pUB307IP501 plasmid was unable to transform PCC7942, indicating that natural transformation of DNA may be restricted by size limitations. The ability to deliver large DNA by conjugation may lead to genetic engineering in PCC7942.

Original languageEnglish
Pages (from-to)15-20
Number of pages6
JournalJournal of Biochemistry
Volume164
Issue number1
DOIs
Publication statusPublished - 2018 Jul 1

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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