Targeted, PCR-based gene disruption in cyanobacteria: inactivation of the polyhydroxyalkanoic acid synthase genes in Synechocystis sp. PCC6803 |
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| Authors: | G Taroncher-Oldenburg G Stephanopoulos |
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| Institution: | (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 |
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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.
Received: 3 March 2000 / Received revision: 1 June 2000 / Accepted: 9 June 2000 |
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