PMID: 2113917Jul 1, 1990Paper

Genetic tagging, cloning, and DNA sequence of the Synechococcus sp. strain PCC 7942 gene (gnd) encoding 6-phosphogluconate dehydrogenase

Journal of Bacteriology
S E Broedel, R E Wolf

Abstract

A genetic approach was used for the cloning of the Synechococcus sp. strain PCC 7942 (Synechococcus strain R2) gnd gene which encodes 6-phosphogluconate dehydrogenase (6PGD). A restriction map of the gnd locus was prepared by Southern analysis using the Escherichia coli gene as a heterologous probe. The Synechococcus strain R2 gene was genetically tagged by restriction site-specific insertion of the nptII gene of Tn903 into a pUC19 plasmid library of Synechococcus strain R2 chromosomal DNA. Synechococcus strain R2 was transformed with this insertion mutation library, and isolates carrying the gnd::nptII gene were identified as mutants hypersensitive to incubation in the dark. The interrupted gene was cloned from one of the mutants. A plasmid carrying the gnd::nptII gene was reintroduced into Synechococcus strain R2, and kanamycin-resistant transformants were selected. Transformants arising by gene replacement were dark sensitive and missing 6PGD activity. Transformants arising by plasmid insertion were dark resistant and had 6PGD activity. The wild-type gene was then cloned from a transformant containing a plasmid insertion, making use of the restriction map derived from the interrupted gene. Synechococcus strain R2 6PGD was ex...Continue Reading

References

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Citations

Oct 29, 2015·Proceedings of the National Academy of Sciences of the United States of America·Benjamin E RubinSusan S Golden
Dec 3, 2016·Proceedings of the National Academy of Sciences of the United States of America·Jared T BroddrickBernhard O Palsson
Mar 14, 2017·Nature Communications·Masahiro KannoShota Atsumi
Jun 9, 2017·PloS One·Gopi Siva Sai Tiruveedula, Pramod P Wangikar
Dec 12, 2018·Trends in Microbiology·David G WelkieSusan S Golden
May 1, 1995·Journal of Bacteriology·D J ScanlanN G Carr
Jan 1, 1993·Molecular and Biochemical Parasitology·M P Barrett, R W Le Page
Sep 10, 2002·Journal of Bacteriology·Olga A Koksharova, C Peter Wolk

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