Isolation and complementation of mutants of Anabaena sp. strain PCC 7120 unable to grow aerobically on dinitrogen
Open Access
- 1 March 1988
- journal article
- research article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 170 (3), 1239-1244
- https://doi.org/10.1128/jb.170.3.1239-1244.1988
Abstract
Mutants of Anabaena sp. strain PCC 7120 unable to grow aerobically on dinitrogen were isolated by mutagenesis with UV irradiation, followed by a period of incubation in yellow light and then by penicillin enrichment. A cosmid vector, pRL25C, containing replicons functional in Escherichia coli and in Anabaena species was constructed. DNA from wild-type Anabaena sp. strain PCC 7120 was partially digested with Sau3AI, and size-fractionated fragments about 40 kilobases (kb) in length were ligated into the phosphatase-treated unique BamHI site of pRL25C. A library of 1,054 cosmid clones was generated in E. coli DH1 bearing helper plasmid pDS4101. A derivative of conjugative plasmid RP-4 was transferred to this library by conjugation, and the library was replicated to lawns of mutant Anabaena strains with defects in the polysaccharide layer of the envelopes of the heterocysts. Mutant EF116 was complemented by five cosmids, three of which were subjected to detailed restriction mapping; a 2.8-kb fragment of DNA derived from one of the cosmids was found to complement EF116. Mutant EF113 was complemented by a single cosmid, which was also restriction mapped, and was shown to be complemented by a 4.8-kb fragment of DNA derived from this cosmid.This publication has 26 references indexed in Scilit:
- Heterocystless mutants ofAnabaenaPCC7120 with nitrogenase activityFEMS Microbiology Letters, 1987
- Nucleotide sequence of the transposon Tn7 gene encoding an aminoglycoside-modifying enzyme, 3″(9)-O-nucleotidyltransferaseNucleic Acids Research, 1985
- Development of the Genetics of Heterocyst-Forming CyanobacteriaCurrent Plant Science and Biotechnology in Agriculture, 1985
- Construction of shuttle vectors capable of conjugative transfer from Escherichia coli to nitrogen-fixing filamentous cyanobacteria.Proceedings of the National Academy of Sciences, 1984
- New Anabaena and Nostoc cyanophages from sewage settling pondsVirology, 1981
- A mutant of Anabaena sp. CA with oxygen-sensitive nitrogenase activityBiochemical and Biophysical Research Communications, 1979
- High recovery of nitrogenase activity and of 55 Fe-labeled nitrogenase in heterocysts isolated from Anabaena variabilisProceedings of the National Academy of Sciences, 1978
- The heterocysts of blue-green algae II. Details of ultrastructureProceedings of the Royal Society of London. B. Biological Sciences, 1971
- Purification of a blue-green algal deoxyribonucleic acid photoreactivating enzyme. An enzyme requiring light as a physical cofactor to perform its catalytic functionBiochemistry, 1970
- New Glycolipid specific to Nitrogen-fixing Blue-green AlgaeNature, 1968