Analysis and Control of Proteolysis of Recombinant Proteins in Escherichia coli

Author(s):  
Aleksei Rozkov ◽  
Sven-Olof Enfors
Author(s):  
Sabreen A Kamal ◽  
Ishraq A Salih ◽  
Hawraa Jawad Kadhim ◽  
Zainab A Tolaifeh

Red rose or roselle (beauty rose ) is natively known as red tea belong to Malvaceae, it is flowers use traditionally for antihypertensive hepato protective, anticancer,antidiabetic,antibacterial, cytotoxicity and antidiarreal, By preparing red tea from it's flower. In this study, we extract chemical compounds by using two solvent which are Ethanol, Ethyl acetate. so we can extract Anthocyanin which is responsible for red colour of flower with many chemical compounds. then study the effect of these extracts on 5 genera from Enterobacteriacaea which can cause diarrheae (Shigella, Salmonella, Escherichia coli, Proteus and Klebsiella ) by preparing 3 concentrations for each solvent (250, 500, 750 ) mg/ml, and control then compare with two antibiotic (Azereonam 30 mg/ml and Bacitracin 10 mg/ml ) these extracts revealed obvious inhibition zone in bacterial growth.


2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Chibuzor M. Nsofor ◽  
Mirabeau Y. Tattfeng ◽  
Chijioke A. Nsofor

Abstract Background This study was aimed to determine the prevalence of qnr genes among fluoroquinolone-resistant Escherichia coli (FREC) isolates from Nigeria. Antimicrobial susceptibility testing was performed by disc diffusion technique. Polymerase chain reaction was used to identify Escherichia coli (E. coli) and for the detection of qnr genes. Results A total of 206 non-duplicate E. coli were isolated from 300 clinical specimens analyzed. In all, 30 (14.6%) of these isolates were FREC; the resistance to fluoroquinolones among these 30 FREC showed 80% (24), 86.7% (26), 86.7% (26), 100% (30), 86.7% (26), 93.3% (28) and 86.7% (26) were resistant to pefloxacin, ciprofloxacin, sparfloxacin, levofloxacin, nalidixic acid, ofloxacin and moxifloxacin, respectively. The distribution of FREC among the various sample sources analyzed showed that 14%, 10%, 13.3%, 16.7% and 20% of the isolates came from urine, stool, high vaginal swab, endo cervical swab and wound swab specimens, respectively. More FREC were isolated from female samples 73.3% (22) compared to male samples 26.7% (8) and were more prevalent among the age group 26–35 years (40%). Twenty eight out of the 30 (93.3%) FREC isolates possessed at least one fluoroquinolone resistance gene in the form of qnrA 10 (33.3%) and qnrB 18 (60%), respectively; qnrS was not detected among the FREC isolates analyzed and 13.5% of the isolates possessed both the qnrA and qnrB genes. Phylogenetic analysis showed that these isolates were genetically diverse. Conclusions These findings suggest a possible resistance to fluoroquinolone is of high interest for better management of patients and control of antimicrobial resistance in Nigeria.


2009 ◽  
Vol 89 (2) ◽  
pp. 285-293 ◽  
Author(s):  
S J Bach ◽  
R P Johnson ◽  
K. Stanford ◽  
T A McAllister

Bacteriophage biocontrol has potential as a means of mitigating the prevalence of Escherichia coli O157:H7 in ruminants. The efficacy of oral administration of bacteriophages for reducing fecal shedding of E. coli O157:H7 by sheep was evaluated using 20 Canadian Arcott rams (50.0 ± 3.0) housed in four rooms (n = 5) in a contained facility. The rams had ad libitum access to drinking water and a pelleted barley-based total mixed ration, delivered once daily. Experimental treatments consisted of administration of E. coli O157:H7 (O157), E. coli O157:H7+bacteriophages (O157+phage), bacteriophages (phage), and control (CON). Oral inoculation of the rams with 109 CFU of a mixture of four nalidixic acid-resistant strains of E. coli O157:H7 was performed on day 0. A mixture of 1010 PFU of bacteriophages P5, P8 and P11 was administered on days -2, -1, 0, 6 and 7. Fecal samples collected on 14 occasions over 21 d were analyzed for E. coli O157:H7, total E. coli, total coliforms and bacteriophages. Sheep in treatment O157+phage shed fewer (P < 0.05) E. coli O157:H7 than did sheep in treatment O157. Populations of total coliforms and total E. coli were similar (P < 0.05) among treatments, implying that bacteriophage lysis of non-target E. coli and coliform bacteria in the gastrointestinal tract did not occur. Bacteriophage numbers declined rapidly over 21 d, which likely reduced the chance of collision between bacteria and bacteriophage. Oral administration of bacteriophages reduced shedding of E. coli O157:H7 by sheep, but a delivery system that would protect bacteriophages during passage through the intestine may increase the effectiveness of this strategy as well as allow phage to be administered in the feed.Key words: Escherichia coli O157:H7, bacteriophage, sheep, environment, coliforms


2007 ◽  
Vol 355 (1) ◽  
pp. 234-239 ◽  
Author(s):  
Tina Schultz ◽  
Jing Liu ◽  
Paola Capasso ◽  
Ario de Marco

2008 ◽  
Vol 74 (23) ◽  
pp. 7431-7433 ◽  
Author(s):  
Mónica Martínez-Alonso ◽  
Nuria González-Montalbán ◽  
Elena García-Fruitós ◽  
Antonio Villaverde

ABSTRACT We have observed that a soluble recombinant green fluorescent protein produced in Escherichia coli occurs in a wide conformational spectrum. This results in differently fluorescent protein fractions in which morphologically diverse soluble aggregates abound. Therefore, the functional quality of soluble versions of aggregation-prone recombinant proteins is defined statistically rather than by the prevalence of a canonical native structure.


2009 ◽  
Vol 13 (12) ◽  
pp. 3274-3284 ◽  
Author(s):  
Kimberly Trabbic-Carlson ◽  
Li Liu ◽  
Bumjoon Kim ◽  
Ashutosh Chilkoti

Biomédica ◽  
2016 ◽  
Vol 36 ◽  
Author(s):  
Ángela Patricia Guerra ◽  
Eliana Patricia Calvo ◽  
Moisés Wasserman ◽  
Jacqueline Chaparro-Olaya

<p><strong>Introducción.</strong> La producción de proteínas recombinantes es fundamental para el estudio funcional de proteínas de <em>Plasmodium</em> <em>falciparum</em>. Sin embargo, las proteínas recombinantes de <em>P</em>. <em>falciparum</em> están entre las más difíciles de expresar y cuando lo hacen usualmente se agregan dentro de cuerpos de inclusión insolubles.</p><p><strong>Objetivo.</strong> Evaluar la producción de cuatro proteínas de <em>P. falciparum</em>, usando como sistema de expresión dos cepas de <em>Escherichia coli </em>genéticamente modificadas para favorecer la producción de proteínas heterólogas y establecer una reserva de proteínas recombinantes puras y solubles y producir anticuerpos policlonales a partir de ellas.<strong></strong></p><p><strong>Materiales y métodos.</strong> Las proteínas recombinantes, las cuales correspondían a secuencias parciales de PfMyoA (Miosina-A) y PfGAP50 (proteína-asociada a glideosoma-50 kDa) y a las secuencias completas de PfMTIP (proteína de interacción con Miosina-A) y PfGAP45 (proteína asociada a glideosoma-45 kDa), fueron expresadas como proteínas de fusión con GST y luego purificadas y usadas para producir anticuerpos policlonales en ratón.</p><p><strong>Resultados.</strong> La expresión de las proteínas recombinantes fue mucho más eficiente en la cepa BL21-CodonPlus (la cual expresa tRNAs escasos en las bacterias silvestres), que en la cepa BL21-pG-KJE8. En contraste, aunque la cepa BL21-pG-KJE sobreexpresa chaperonas, no redujo la formación de cuerpos de inclusión. <strong>Conclusión.</strong> El uso de cepas de <em>E</em>. <em>coli</em> genéticamente modificadas fue fundamental para alcanzar altos niveles de expresión de las cuatro proteínas recombinantes evaluadas y permitió obtener dos de ellas en forma soluble. La estrategia utilizada permitió expresar cuatro proteínas recombinantes de <em>P</em>. <em>falciparum</em> en cantidad suficiente para inmunizar ratones y producir anticuerpos policlonales, y además conservar proteína pura y soluble de dos de ellas, para ensayos futuros.</p>


2009 ◽  
Vol 25 ◽  
pp. S244
Author(s):  
Y. Sevastsyanovich ◽  
S. Alfasi ◽  
L. Griffiths ◽  
T. Overton ◽  
J. Cole

1991 ◽  
Vol 646 (1 Recombinant D) ◽  
pp. 254-258 ◽  
Author(s):  
DIOGO ARDAILLON SIMOES ◽  
MALENE DAL JENSEN ◽  
ERIC DREVETON ◽  
MARIE-ODILE LORET ◽  
SYLVIE BLANCHIN-ROLAND ◽  
...  

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