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Targeted, PCR-based gene disruption in cyanobacteria: inactivation of the polyhydroxyalkanoic acid synthase genes in Synechocystis sp. PCC6803

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Abstract

A PCR-based method is described for the efficient construction of targeted gene disruptions and gene fusions in the cyanobacterium Synechocystis sp. PCC6803. In a simple two-step PCR approach, a gene conversion cassette was synthesized targeting the polyhydroxyalkanoic acid (PHA) synthase genes. Upon transformation, PHA production in Synechocystis under normal as well as high production culture conditions was undetectable. The application of this method to the genetic inactivation of the phaE-C Syn gene cluster demonstrates its potential for genetic engineering of cyanobacteria and the study of functional genomics in Synechocystis.

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Authors and Affiliations

  1. Massachusetts Institute of Technology, Chemical Engineering Department, MIT 56–422, 77 Massachusetts Ave., Cambridge, MA 02139, USA e-mail: gto@mit.edu Tel.: +1-617-2580349 Fax: +1-617-2533122, , , , , , US

    G. Taroncher-Oldenburg & G. Stephanopoulos

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  1. G. Taroncher-Oldenburg
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  2. G. Stephanopoulos
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Received: 3 March 2000 / Received revision: 1 June 2000 / Accepted: 9 June 2000

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Taroncher-Oldenburg, G., Stephanopoulos, G. Targeted, PCR-based gene disruption in cyanobacteria: inactivation of the polyhydroxyalkanoic acid synthase genes in Synechocystis sp. PCC6803. Appl Microbiol Biotechnol 54, 677–680 (2000). https://doi.org/10.1007/s002530000450

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  • DOI: https://doi.org/10.1007/s002530000450

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