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PMID: 193099 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Gene cloning for the isolation of enzymes of membrane lipid synthesis: phosphatidylserine synthase overproduction in Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 74 ·No. 4 ·1977-04-00 ·Pages 1412-6

Raetz CR, Larson TJ, Dowhan W

Abstract

We have screened a bank of 2000 E. coli strains carrying hybrid ColE1 plasmids [Clarke, L. & Carbon, J. (1976) Cell 9, 91-99] for those that correct the temperature sensitivity of a mutant in CDP-1,2-diacyl sn-glycerol:L-serine O-phosphatidyltransferase (EC 2.7.8.8, phosphatidylserine synthase). Two hybrid plasmids of this kind (pLC34-44 and pLC34-46) were identified and characterized. Strains carrying these plasmids overproduce the synthase by 6- to 15-fold, as demonstrated by assays of extracts and purification to homogeneity of the overproduced enzyme. The overproduced synthase, like the wild-type enzyme, is found associated predominately with the ribosomal fraction of crude cell extracts. Because the membrane phospholipid composition of these overproducers is not greatly altered, we suggest that the synthase is normally present in excess.

MeSH Terms
CDPdiacylglycerol-Serine O-Phosphatidyltransferase/biosynthesis Escherichia coli/enzymology,metabolism Extrachromosomal Inheritance Genes Genotype Hybridization, Genetic Mutation Phospholipids/metabolism Phosphotransferases/biosynthesis Plasmids Protein Biosynthesis Species Specificity Subcellular Fractions/enzymology Transcription, Genetic
Chemicals
Phospholipids Phosphotransferases CDPdiacylglycerol-Serine O-Phosphatidyltransferase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Raetz C R
Larson T J
Dowhan W
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21 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.74.4.1412
Published
1977-04-00
Pages
1412-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC430779
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.74.4.1412 PMC Full Text View journal details
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