Abstract:
:Bacteriophage T7 RNA polymerase is stable in Escherichia coli but very susceptible to cleavage by at least one endoprotease after cell lysis. The major source of this endoprotease activity was found to be localized to the outer membrane of the cell. A rapid whole-cell assay was developed to screen different strains for the presence of this proteolytic activity. Using this assay, we identified some common laboratory strains that totally lack the protease. Genetic and Southern analyses of these null strains allowed us to conclude that the protease that cleaves T7 RNA polymerase is OmpT (formerly termed protein a), a known outer membrane endoprotease, and that the null phenotype results from deletion of the OmpT structural gene. A recombinant plasmid carrying the ompT gene enables these deletion strains to synthesize OmpT and converts them to a protease-positive phenotype. The plasmid led to overproduction of OmpT protein and protease activity in the E. coli K-12 and B strains we used, but only weak expression in the E. coli C strain, C1757. This strain-dependent difference in ompT expression was investigated with respect to the known influence of envZ on OmpT synthesis. A small deletion in the ompT region of the plasmid greatly diminishes the amount of OmpT protein and plasmid-encoded protease present in outer membranes. Use of ompT deletion strains for production of T7 RNA polymerase from the cloned gene has made purification of intact T7 RNA polymerase routine. Such strains may be useful for purification of other proteins expressed in E. coli.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Grodberg J,Dunn JJdoi
10.1128/jb.170.3.1245-1253.1988subject
Has Abstractpub_date
1988-03-01 00:00:00pages
1245-53issue
3eissn
0021-9193issn
1098-5530journal_volume
170pub_type
杂志文章abstract::The enzymatic properties and the physiological function of the type IV apurinic/apyrimidinic (AP)-endonuclease homolog of Bacillus subtilis, encoded by yqfS, a gene specifically expressed in spores, were studied here. To this end, a recombinant YqfS protein containing an N-terminal His6 tag was synthesized in Escheric...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.185.18.5380-5390.2003
更新日期:2003-09-01 00:00:00
abstract::Spiralin could not be solubilized in the absence of detergents, and it was shown by charge-shift crossed immunoelectrophoresis that this protein was capable of binding detergents under nondenaturing conditions. These properties indicate the amphiphilic nature of spiralin, which therefore should be regarded as an intri...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.140.2.738-741.1979
更新日期:1979-11-01 00:00:00
abstract::Purified bulk tRNA from Methanococcus vanielii (carbon source, formate) showed variation in the modified nucleoside pattern reported for Escherichia coli as analyzed by both ion-exchange and thin-layer chromatography. Ribothymidine and 7-methylguanosine were absent; 1-methyladenosine, 1-methylguanosine, N2-methylguano...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.133.1.240-250.1978
更新日期:1978-01-01 00:00:00
abstract::The cellular fatty acid composition of Mycobacterium vaccae JOB5 and Mycobacterium convolutum R22 was examined after growth on n-alkanes and compared with the fatty acids of the organisms after growth on 1-chlorohexadecane and 1-chlorooctadecane. Growth on n-alkanes resulted in normal fatty acid profiles. Mass spectra...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.156.3.1158-1164.1983
更新日期:1983-12-01 00:00:00
abstract::We previously described mutations in a gene, secB, which have pleiotropic effects on protein export in Escherichia coli. In this paper, we report the isolation of mutants in which the activity of the secB gene was eliminated. Null mutations in secB affected only a subset of exported proteins. Strains carrying these mu...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.163.1.267-274.1985
更新日期:1985-07-01 00:00:00
abstract::The stable inheritance of the broad-host-range plasmid RK2 is due at least in part to functions within a region located at coordinates 32.8 to 35.9 kb, termed the RK2 par locus. This locus encodes four previously identified genes in two operons (parCBA and parD; M. Gerlitz, O. Hrabak, and H. Schwab, J. Bacteriol. 172:...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.174.24.8119-8132.1992
更新日期:1992-12-01 00:00:00
abstract::Mutations in carbohydrate-negative mutants of Pseudomonas aeruginosa PAO1 individually deficient in glucose 6-phosphate dehydrogenase (zwf), 6-phosphogluconate dehydratase (edd), or pyruvate carboxylase (pyc) were mapped on the chromosome by plasmid R68.45-mediated conjugation and by bacteriophage F116L-mediated trans...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.156.3.1123-1129.1983
更新日期:1983-12-01 00:00:00
abstract::Streptococcus faecalis JH1 contains two conjugative plasmids, pJH1, an R plasmid that codes for resistance to kanamycin, streptomycin, erythromycin, and tetracycline, and pJH2, a hemolysin-bacteriocin plasmid. Strain JH1 was used as an antibiotic resistance donor in conjugation experiments with two plasmid-free S. fae...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.155.3.1094-1104.1983
更新日期:1983-09-01 00:00:00
abstract::Sigma 54-dependent transcriptional activators such as Escherichia coli NtrC, Rhizobium meliloti NifA, and Rhizobium leguminosarum DctD share similar central and carboxy-terminal domains but differ in the structure and function of their amino-terminal domains. We have deleted the amino-terminal and carboxy-terminal dom...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.174.4.1428-1431.1992
更新日期:1992-02-01 00:00:00
abstract::Spore cortex of conditional cortexless mutants of Bacillus sphaericus 9602 was not detectable by electron microscopy unless the medium was supplemented with meso-alpha,epsilon-diaminopimelic acid during sporulation. Other spore structures appeared normal. Spore shape was quite irregular in the absence of meso-alpha,ep...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.127.3.1568-1570.1976
更新日期:1976-09-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.184.4.1003-1009.2002
更新日期:2002-02-01 00:00:00
abstract::In Salmonella typhimurium the two enzymes of proline catabolism, proline oxidase and Delta(1)-pyrroline-5-carboxylic acid dehydrogenase, are subject to catabolite repression when the cells are grown in the presence of glucose. Mutants partially relieved of catabolite repression (PutR) for the proline catabolic enzymes...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.111.2.375-382.1972
更新日期:1972-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00439-07
更新日期:2007-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00674-16
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.179.5.1609-1613.1997
更新日期:1997-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.169.8.3450-3457.1987
更新日期:1987-08-01 00:00:00
abstract::Many microbes coexist within biofilms, or multispecies communities of cells encased in an extracellular matrix. However, little is known about the microbe-microbe interactions relevant for creating these structures. In this study, we explored a striking dual-species biofilm between Bacillus subtilis and Pantoea agglom...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00670-18
更新日期:2019-08-22 00:00:00
abstract::To investigate the involvement of bacterial catalases of the symbiotic gram-negative bacterium Rhizobium meliloti in the development of Medicago-Rhizobium functional nodules, we cloned a putative kat gene by screening a cosmid library with a catalase-specific DNA probe amplified by PCR from the R. meliloti genome. Nuc...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.178.23.6802-6809.1996
更新日期:1996-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.126.2.997-998.1976
更新日期:1976-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.175.8.2423-2435.1993
更新日期:1993-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.10.4890-4896.1988
更新日期:1988-10-01 00:00:00
abstract::Inorganic phosphate was found to decrease the caseinolytic and ethyl-esterase activities of the Microsporum gypseum germination protease. The germination protease possessed exokeratinase (beta-keratinase) activity immediately after release from the fungal spore. After phosphate treatment of the enzyme, the germination...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.117.2.422-431.1974
更新日期:1974-02-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.1.393-399.1988
更新日期:1988-01-01 00:00:00
abstract::Salmonella typhimurium and Escherichia coli from anaerobic cultures displayed tactic responses to gradients of nitrate, fumarate, and oxygen when the appropriate electron transport pathway was present. Such responses were named "electron acceptor taxis" because they are elicited by terminal electron acceptors. Mutant ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.140.2.567-573.1979
更新日期:1979-11-01 00:00:00
abstract::Streptococcus suis serotype 2 is a highly invasive, extracellular pathogen in pigs with the capacity to cause severe infections in humans. This study was initiated by the finding that IgM degradation products are released after opsonization of S. suis. The objective of this work was to identify the bacterial factor re...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01875-12
更新日期:2013-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.180.3.647-654.1998
更新日期:1998-02-01 00:00:00
abstract::The delta subunit of RNA polymerase, RpoE, is widespread in low-G+C Gram-positive bacteria and is thought to play a role in enhancing transcriptional specificity by blocking RNA polymerase binding at weak promoter sites and stimulating RNA synthesis by accelerating core enzyme recycling. Despite the well-studied bioch...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00653-10
更新日期:2010-10-01 00:00:00
abstract::The expression of the pur operon, which encodes enzymes of the purine biosynthetic pathway in Bacillus subtilis, is subject to control by the purR gene product (PurR) and phosphoribosylpyrophosphate. This control is also exerted on the purA and purR genes. A consensus sequence for the binding of PurR, named the PurBox...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.183.21.6175-6183.2001
更新日期:2001-11-01 00:00:00
abstract::A large-scale fractionation scheme purified the major molybdenum(Mo)-binding protein (Mop) from crude extracts of Clostridium pasteurianum, with a 10 and 0.2% yield of Mo and protein, respectively. The apparent molecular weight of the purified molybdoprotein is 5,700, as determined by sodium dodecyl sulfate-polyacryla...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.168.2.688-693.1986
更新日期:1986-11-01 00:00:00
abstract::Evolutionary trajectories and mutational landscapes of drug-resistant bacteria are influenced by cell-intrinsic and extrinsic factors. In this study, I demonstrated that loss of the Lon protease altered susceptibility of Escherichia coli to trimethoprim and that these effects were strongly contingent on the drug conce...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00561-19
更新日期:2020-01-15 00:00:00