Effect of isopropyl-beta-D-thiogalactopyranosid induction of the lac operon on the specificity of spontaneous and doxorubicin-induced mutations in Escherichia coli.
'Previous studies of doxorubicin-induced mutations employing F'' lacl/lacO as an endogenous gene target have focused on properties of large deletions with 3'' endpoints residing in the lacO region of the target gene. This study considers the influence of Lac repressor binding on the distribution of these deletions. Results of the ... More
New M13 vectors for cloning.
AuthorsMessing J
JournalMethods Enzymol
PubMed ID6310323
Substrate-induced inactivation of a crippled beta-glucosidase mutant: identification of the labeled amino acid and mutagenic analysis of its role.
AuthorsGebler JC, Trimbur DE, Warren AJ, Aebersold R, Namchuk M, Withers SG
JournalBiochemistry
PubMed ID7578060
The beta-glucosidase from Agrobacterium sp. catalyzes the hydrolysis of beta-glucosides via a covalent alpha-D-glucopyranosyl-enzyme intermediate involving Glu358. Hydrolysis of 2,4-dinitrophenyl beta-D-glucopyranoside by the low activity Glu358Asp mutant of Agrobacterium beta-glucosidase is accompanied by time-dependent inactivation of the enzyme. Through kinetic studies, labeling, and sequence analysis, inactivation is shown to be ... More
Flow cytometry sorting of viable bacteria and yeasts according to beta-galactosidase activity.
AuthorsNir R, Yisraeli Y, Lamed R, Sahar E
JournalAppl Environ Microbiol
PubMed ID2128011
We describe a novel method for quantitative measurement of beta-galactosidase (beta-gal) levels in bacteria and yeasts by using flow cytometry, a method which allows viable microbial cells to be sorted on the basis of the expressed activity and to be recultivated. The method is based on encapsulating single cells in ... More
Cell-division control in Escherichia coli: specific induction of the SOS function SfiA protein is sufficient to block septation.
AuthorsHuisman O, D'Ari R, Gottesman S
JournalProc Natl Acad Sci U S A
PubMed ID6087326
Blocks in DNA replication cause a rapid arrest of cell division in Escherichia coli. We have previously established that the function SfiA (SulA), induced under these conditions as part of the SOS response, is involved in this inhibition of division. To separate the effects of SfiA from those of other ... More
Membrane topology of helices VII and XI in the lactose permease of Escherichia coli studied by lacY-phoA fusion analysis and site-directed spectroscopy.
The use of lactose permease-alkaline phosphatase fusions (lacY-phoA) demonstrates that the lactose permease of Escherichia coli contains 12 transmembrane domains and that approximately half of a transmembrane domain is required to translocate alkaline phosphatase to the periplasmic surface of the membrane [Calamia, J., & Manoil, C. (1990) Proc. Natl. Acad. ... More
Expression of human liver fatty acid-binding protein in Escherichia coli and comparative analysis of its binding characteristics with muscle fatty acid-binding protein.
AuthorsMaatman RG, van Moerkerk HT, Nooren IM, van Zoelen EJ, Veerkamp JH
JournalBiochim Biophys Acta
PubMed ID8068722
Human liver fatty acid-binding protein (L-FABP) has been efficiently expressed in Escherichia coli. The cDNA encoding human liver FABP was under the control of T7 RNA polymerase promoter in the expression vector pET-3b. Expression required overnight induction with isopropyl beta-D-thiogalactopyranoside in the presence of the bacterial RNA polymerase inhibitor, rifampicin. ... More