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Characterization and mode of action of a bacteriocin produced by a Bacteroides Fragilis strainMossie, Godwin Mxolisi Kevin January 1980 (has links)
Bacteroides fragilis strain Bf-1 produces an extracellular bacteriocin at the beginning of the stationary growth phase. Production is not inducible by either ultraviolet light or mitomycin C. The low molecular weight bacteriocin (MW estimates of 13 500 and 18 800 obtained from Sephadex G-100 chromatography and SDS-PAGE electrophoresis respecively) is stable between pH 7 - 9 and is inactivated on incubation with trypsin and pronase. An unusual feature of the Bf-1 bacteriocin is its apparent biphasic temperature stability: while the majority of the activity (97%) is destroyed by heating at 60ºC (t [subscript] 1/2 = 2.5 min at 60ºC), a small proportion (3%) is stable even after autoclaving at 121ºC for 15 min. The killing of sensitive cells occurs in 2 stages and the killing action is reversed by incubation with trypsin. The transition from stage I to stage II is dependent on the temperature of incubation and the growth state of sensitive cells. 2,4-Dinitrophenol prevents this transition. The Bf-1 bacteriocin has an unusual mode of action. It specifically inhibits RNA synthesis whilst having no effect on protein or DNA synthesis. No effect on intracellular ATP levels were observed. The heat-stable (3%) fraction had a similar biochemical effect. In vitro studies involving RNA polymerase indicated that the bacteriocin and the antibiotic rifampicin have similar effects on RNA synthesis. The bacteriocinogenic strain (Bf-1) is insensitive to its own bacteriocin both in vivo and in vitro, although this immunity is overcome in vitro by the addition of higher concentrations of the Bf-1 bacteriocin. The bacteriocinogenic strain (Bf-1) harbors a cryptic plasmid (or plasmids) which on a neutral sucrose gradient, sediments faster than the Col E1 marker plasmid DNA. Attempts to cure this strain of its bacteriocinogenic phenotype were unsuccessful.
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Protein synthesis in the ovine fetusSchaefer, Allan Lee January 1983 (has links)
To study the effect of maternal nutrition on fetal protein
synthesis, isotopic kinetic studies were undertaken in which
indwelling catheters were implanted in the inferior vena cava
and saphenous vein of ovine fetuses as well as the jugular vein
and femoral artery of ewes at 120-130 days of gestation
Following a five day post surgical recovery period an 8h
continuous infusion of L-[2,3,5,6-³H] or L-[U-¹⁴ C] tyrosine was made into the fetus and ewe simultaneously. An enzymatic procedure specific for L-tyrosine was used to measure the plateau specific activity in fetal and maternal plasma. Using these values the net placental transfer, endogenous production and net utilization of tyrosine by the fetus were determined employing a two pool kinetic model. For determining the fractional protein synthetic rates in individual tissues the ewes were sacrificed and the fetuses obtained by laparotomy immediately following the continuous infusion of labelled tyrosine. The effect of starvation on tyrosine transfer across the placenta and protein synthesis by the fetus was studied by repeating the above procedures in ewes starved for 48h. Values for the net placental transfer, endogenous production and net utilization of tyrosine were 5.14, 1.29 and 6.42 mmol/d/kg respectively for fetuses of fed ewes, whereas, these values were -0.29, 4.53 and 3.88 mmol/d/kg in fetuses of starved ewes. The plasma concentrations (mg%) of glucose, alpha amino nitrogen and lactate in the fed ewes were 60.18, 10.68 and 5.58 versus 43.66, 7.80 and 7.41 in starved ewes. Glucose, alpha amino nitrogen and lactate values were also seen to change in the fetus from 12.55,
13.38 and 12.98, respectively in the fed versus 8.18, 11.55 and 15.08 in fetuses of starved ewes. The fetal tyrosine net utilization values, corrected for oxidative loss of 5.2%, were used to calculate the whole body protein synthetic values. From an average tyrosine content of carcass protein of 15.71 mmol/kg, the whole body protein synthetic rates were 63 g/d/kg in the fed versus 25 g/d/kg in the starved fetuses. Fetal tissue fractional protein synthetic rates were also affected by maternal starvation. The fractional synthetic rates (%/day) for liver, kidney, lung, heart and skeletal muscle were 78%, 45%, 65%, 14% and 26% for fed versus 12%, 22%, 23%, 11% and 10%/d for the fetuses of starved ewes. The data demonstrate that maternal feed deprivation reduces protein synthesis in the fetus. / Land and Food Systems, Faculty of / Graduate
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Characterization of immediate-early and early proteins of murine cytomegalovirus synthesized in permissive and nonpermissive cellsWalker, Douglas Gordon January 1985 (has links)
The gene products produced by murine cytomegalovirus (MCMV) in infected cells prior to viral DNA synthesis are believed to control the interaction of the virus with the cells, determining whether a permissive infection results, with virus replication, or whether further virus gene expression is inhibited, resulting in a latent or abortive infection. The aim of this study was to characterize the early viral gene products that are produced in permissive and nonpermissive cells.
The proteins produced in 3T3-L1 cells, permissively infected with MCMV, during the first six hours of infection (the period prior to viral DNA replication) were characterized by polyacrylamide gel electrophoresis. Ten of the proteins were classified as immediate-early (IE) and seven as early according to their time of synthesis and also according to their synthesis in the presence of actinomycin D following the reversal of a cycloheximide mediated block in protein synthesis. The estimated molecular weights ranged from 28K - 100K. The synthesis of a dominant IE protein of 100K was significantly increased, after the reversal of a cycloheximide block, compared to unenhanced conditions. The synthesis of two other major IE proteins of 96K and 89K were also significantly enhanced by this treatment. The 100K and 89K proteins partitioned with the nuclear, cytoplasmic and cytoskeletal fractions, while the 96K protein partitioned more strongly with the nuclei. These proteins were phosphorylated. The other IE proteins were synthesized in lesser amounts. The major early proteins, which had molecular weights of 39K and 36K, were also phosphorylated and were exclusively nucleus-associated. A number of the IE and early proteins had affinity for native and denatured DNA-cellulose.
The same major IE and early proteins were identified in nonpermissively infected J774A.1 macrophage cells. Although 0.6% of these cells became permissively infected with MCMV and the rest appeared to be nonpermissively infected, viral DNA and late protein synthesis was not detected. The major difference between the proteins produced in 3T3-L1 cells and J774A.1 cells was the affinity of the 96K protein for denatured DNA-cellulose, which was only observed when the protein was synthesized in J774A.1 cells.
The main IE and early MCMV induced proteins were also synthesized in nonpermissively infected human fibroblast cells. The only difference between the proteins produced in these cells and 3T3-L1 cells was that the 100K IE protein appeared to have a greater nuclear-affinity, when produced in the human fibroblasts, than was found when synthesized in infected 3T3-L1 cells.
In conclusion, a larger number of IE and early MCMV-induced proteins were identified in infected cells than had been previously characterized. There was no evidence of restricted MCMV gene expression occurring in two different cell types that were nonpermissively infected. This appeared to indicate that, in the nonpermissive experiments described, MCMV replication was inhibited at the stage of viral DNA synthesis. / Medicine, Faculty of / Pathology and Laboratory Medicine, Department of / Graduate
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Basic nuclear protein synthesis during the cell cycle analysed by unit gravity sedimentation.Tang, Shun Chii. January 1972 (has links)
No description available.
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An electron microscope study of organelle autonomy in Ochromonas danica using inhibitors of protein synthesis.Smith-Johannsen, Heidi January 1971 (has links)
Note:
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Protein synthesis and drought stress in two rapeseed cultivarsLeblanc, Rosanne January 1991 (has links)
No description available.
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Development of Methylobacterium extorquens as a recombinant protein production system and the expression of the heterologous cry1Aa gene from Bacillus thuringiensisBélanger, Louise January 2003 (has links)
No description available.
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Protein synthesis in a piglet model of gastrointestinal inflammation and malnutritionMackenzie, Michelle Lee. January 2001 (has links)
No description available.
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Cell free synthesis of ferritin using the Modified Reticulocyte Lysate System.Clark, Nathaniel Goodwin. January 1978 (has links)
Thesis: M.S., Massachusetts Institute of Technology, Department of Nutrition and Food Science, 1978 / Includes bibliographical references. / M.S. / M.S. Massachusetts Institute of Technology, Department of Nutrition and Food Science
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Investigation of Mycobacterium tuberculosis protein expression and analysis of humoral immune responses of TB patientsPheiffer, Carmen 12 1900 (has links)
Thesis (PhD)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: New agents for the diagnosis, prevention and treatment of tuberculosis are urgently
required. Yet, despite extensive tuberculosis research over recent years, no new drugs,
vaccines or diagnostics have been identified to date. It is widely speculated that the major
obstacle to the identification of new therapies is the lack of understanding of the hostpathogen
interaction.
This study has investigated whether patterns of antigen expression correlate with
molecular epidemiological data and strain virulence through the analysis of protein expression
and antigen recognition profiles of different M tuberculosis clinical isolates. Using
polyacrylamide gel electrophoresis, enzyme-linked immunosorbent assay, and Western
blotting, protein expression and antigen recognition by two genotypically different clinical
strains that differed in their frequency in the study population have been compared. In
addition to differences in protein expression and antigen recognition between the clinical
strains and the reference strain H37Rv, protein expression differences between the clinical
strains themselves were observed which may relate to strain frequency and virulence.
Differential protein expression by M tuberculosis strains, may explain the
heterogeneous host humoral immune response and why no fully effective serodiagnostic test
has been developed to date. To explore this hypothesis, the potential of serodiagnosis in this
community, where patients are infected with a wide variety of genotypically distinct strains,
was investigated. IgG levels to three mycobacterial antigens showed that serodiagnosis of TB
is possible in this community, despite infection by a wide variety of genotypically different
M tuberculosis strains. Disease episode affected antibody levels, suggesting that care should
be taken when evaluating serological diagnosis for repeat episode patients. This study has shown that M tuberculosis protein expression is dynamic and that the
bacillus presents a hypervariabie array of antigens to the host immune system. It is likely that
different antigens become immunodominant as antituberculosis chemotherapy progresses, and
that these differentially expressed antigens may be tracked as predictors of treatment outcome.
This hypothesis was tested by correlating Ag85-specific IgG with treatment response, as
assessed by sputum smear conversion after two months of antimycobacterial chemotherapy.
No significant correlation between antibody levels and treatment responses was observed,
suggesting that antibodies may not be useful surrogate markers or that the incorrect antibody
type or mycobacterial antigen were selected. Results were consistent with previous findings
where patient-to-patient variation dictated the host humoral response.
The results obtained in this study have demonstrated that although bacteriological
factors may influence strain prevalence due to antigen variation and immune evasion, both
bacteriological and host factors affect humoral immunity. Differential protein expression by
M tuberculosis strains has potentially important implications for serodiagnosis and the
development of subunit or DNA vaccines, by suggesting that multi-antigen cocktails should
be used. Differential protein expression may also explain why patients do not develop
adequate protective immunity and are susceptible to reinfection. / AFRIKAANSE OPSOMMING: Daar is 'n dringende behoefte vir nuwe middels vir die diagnosering, voorkoming en
behandeling van tuberkulose. Ondanks intense tuberkulose navorsing gedurende die afgelope
paar jaar, is daar geen nuwe tuberkulose medikasie, vaksines of diagnostiese metodes
geïdentifiseer nie. Daar word gespekuleer dat die hoof struikelblok vir die identifisering van
nuwe medikasie die onkunde oor die tuberkulose patogeen is.
Deur die analise van proteien-uitdrukking en antigeen-erkenning profiele van
verskillende M. tuberculosis kliniese isolate is daar tydens hierdie studie ondersoek ingestel
of die patroon van antigeen uitdrukking korreleer met molekulêre epidemiologiese data and
stam-virulensie. Proteien-uitdrukking en antigeen-erkenning deur twee genotipies
verskillende kliniese stamme wat verskil in hul frekwensie in die bestudeerde populasie, is
vergelyk deur middel van poli-akrielamied gel elektroforese, ensiem-gekoppelde
immuunabsorberende analise en Westelike oordrag. Addisoneel tot die verskille in proteienuitdrukking
en antigeen-ekenning tussen kliniese stamme en die verwysingstam H37Rv, is
daar ook verskille aangedui tussen die kliniese stamme self wat kan dui op stam frekwensie
en virulensie.
Differensiële proteien-uitdrukking deur M. tuberculosis stamme, kan moontlik die
heterogene gasheer se humorale immuunreaksie verduidelik en daarmee saam die rede
waarom daar nie tot op hede 'n effektiewe sero-diagnostiese toets ontwikkel is nie. Daar is
dus ondersoek ingestel na die potensiaal van sero-diagnose in 'n gemeenskap waar pasiënte
geïnfekteer is met 'n wye verskeidenheid genotipiese stamme. Die IgG vlakke van drie
mikobakteriële antigene het aangedui dat sero-diagnose van tuberkulose moontlik is in hierdie
gemeenskap, ten spyte van infektering deur 'n wye verskeidenheid genotipies-verskillende M.
tuberculosis stamme. Die tussenspel van die siekte het teenliggaampie-vlakke beïnvloed wat daarop dui dat daar versigtig moet gelet word tydens die evaluering van serologiese diagnose
van geïnfekteerde pasiënte wat voorheen siek was.
Hierdie studie toon dat M. tuberculosis proteïen-uitdrukking dinamies is en dat die
bacillus 'n groot variëteit van antigene tot die immuun sisteem bied. Dit is moontlik dat
verskillende antigene immuun dominant kan word soos wat antituberkulose chemoterapie
toeneem, en dat hierdie verskillend-uitgedrukte antigene as 'n gevolg daarvan gebruik kan
word as voorspellers vir behandeling. Hierdie hipotese is getoets deur die korrelering van
Ag85-spesifieke IgG met die reaksie op behandeling soos geëvalueer deur speeksel-monster
verandering na twee maande se anti-mikobakteriële chemoterapie. Daar was geen
noemenswaardige korrelasie tussen teenliggaampie vlakke en die reaksie op behandeling nie,
wat daarop dui dat die teenliggaampies nie toepaslike surrogaat merkers is nie of dat die
verkeerde teenliggaampie-tipe of mikobakteriële antigeen geselekteer is. Hierdie resultate
bevestig vorige bevindinge waar pasiënt-tot-pasiënt verskille die gasheer se humorale
immuunreaksie gedikteer het.
Die resultate wat uit hierdie studie volg dui dat alhoewel bakteriologiese faktore die
stam-frekwensie kan beïnvloed as gevolg van antigeen-variasie en immuun-ontduiking, kan
beide bakteriologiese en gasheer faktore die humorale immuunreaksie beïnvloed.
Differensiële proteiën uitdrukking deur 'n verskeidenheid M. tuberculosis stamme het
potensieël belangrike toepassings vir sero-diagnose en die ontwikkeling van subeenheid of
DNS vaksines wat impliseer dat multi-antigeen mengsels gebruik moet word. Differensiële
proteiën uitdrukking mag ook verduidelik waarom pasiënte nie 'n voldoende beskermende
immuniteit opbou nie en sodoende ontvanklik is vir her-infeksie.
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