PMID: 49056Jun 1, 1975Paper

Isolation of Escherichia coli mutants defective in enzymes of membrane lipid synthesis

Proceedings of the National Academy of Sciences of the United States of America
C R Raetz

Abstract

A new method has been developed which permits the rapid screening of E. coli colonies for mutants with defective enzymes of phospholipid metabolism. In this procedure, a disc of filter paper is pressed down on an agar plate containing several hundred colonies of mutagen-treated cells, after which the paper is lifted off. In the process the colonies are transferred to the paper, giving rise to a replica print of the master plate. The few cells from each colony left on the master keep growing in the original pattern. The pattern of colonies is also retained on the filter paper, even after the cells are rendered permeable with lysozyme and EDTA. Colonies treated in this manner remain absorbed to the paper, where they can convert sn-(U-14-C)glycero-3-P to phosphatidyl(U-14-C)glycerophosphate, dependent on added CDP-diglyceride. Unrelated reactions of sn-(U-14-C)glycero-3-P that may obscure the synthesis of phosphatidyl-glycerophosphate are inhibited by the addition of reagents poisoning energy generation. The radioactive phospholipid that forms around each colony on the paper is precipitated in situ with trichloroacetic acid, and unreacted sn-(U-14-C)glycero-3-P is washed away. After autoradiography, the colonies on the filter pape...Continue Reading

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Citations

Dec 12, 2012·World Journal of Microbiology & Biotechnology·Md Rezaul BariNobuhiro Mori
Mar 21, 1986·Biochimica Et Biophysica Acta·T H Chiu, L L Yeh
Jul 9, 1999·Progress in Lipid Research·C C DiRussoJ D Weimar
Mar 11, 2000·European Journal of Biochemistry·S KochharJ E Germond
Jan 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·T Y Chang, P R Vagelos
Feb 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·P K Mukherjee, H Paulus
Apr 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·C R RaetzW Dowhan
Mar 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·J D Esko, C R Raetz
Apr 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·A R Robbins
Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·U HenningC P Hollenberg
Sep 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·J D Esko, C R Raetz
Nov 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·C MiyazakiI Shibuya
Jan 2, 1987·European Journal of Biochemistry·K Hosaka, S Yamashita
Mar 20, 2012·Applied and Environmental Microbiology·Matthew D McMahonMichael G Thomas
Oct 25, 2007·Proceedings of the National Academy of Sciences of the United States of America·Nick Zagorski
Mar 12, 2013·Annual Review of Biochemistry·William DowhanPaul Modrich
Aug 1, 1979·Analytical Biochemistry·U HenningR Chen
Aug 11, 2004·Analytical Biochemistry·Christoph Benning
Nov 18, 2004·Biochimica Et Biophysica Acta·Annemieke van Dalen, Ben de Kruijff
May 12, 2017·The Journal of Biological Chemistry·William Dowhan
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Jan 27, 2021·Proceedings of the National Academy of Sciences of the United States of America·Reed B Wickner
Apr 13, 2021·Frontiers in Molecular Biosciences·William Dowhan, Mikhail Bogdanov
May 1, 1982·Journal of Bacteriology·J NikawaS Yamashita

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