scholarly journals Purification of recombinant proteins from Escherichia coli at low expression levels by inverse transition cycling

2007 ◽  
Vol 360 (1) ◽  
pp. 166-168 ◽  
Author(s):  
Trine Christensen ◽  
Kimberly Trabbic-Carlson ◽  
Wenge Liu ◽  
Ashutosh Chilkoti
Keyword(s):  
Escherichia Coli ◽  
Recombinant Proteins ◽  
Expression Levels ◽  
Inverse Transition Cycling ◽  
Inverse Transition ◽  
Low Expression

scholarly journals Low expression of NSD1, NSD2, and NSD3 define a subset of human papillomavirus-positive oral squamous carcinomas with unfavorable prognosis

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Steven F. Gameiro ◽  
Farhad Ghasemi ◽  
Peter Y. F. Zeng ◽  
Neil Mundi ◽  
Christopher J. Howlett ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
T Cell ◽  
The Cancer Genome Atlas ◽  
Cell Infiltration ◽  
Relative Level ◽  
Expression Levels ◽  
T Cell Infiltration ◽  
Molecular Features ◽  
Platinum Based Chemotherapy ◽  
Low Expression

Abstract Background Frequent mutations in the nuclear receptor binding SET domain protein 1 (NSD1) gene have been observed in head and neck squamous cell carcinomas (HNSCC). NSD1 encodes a histone 3 lysine-36 methyltransferase. NSD1 mutations are correlated with improved clinical outcomes and increased sensitivity to platinum-based chemotherapy agents in human papillomavirus-negative (HPV-) tumors, despite weak T-cell infiltration. However, the role of NSD1 and related family members NSD2 and NSD3 in human papillomavirus-positive (HPV+) HNSCC is unclear. Methods Using data from over 500 HNSCC patients from The Cancer Genome Atlas (TCGA), we compared the relative level of mRNA expression of NSD1, NSD2, and NSD3 in HPV+ and HPV- HNSCC. Correlation analyses were performed between T-cell infiltration and the relative level of expression of NSD1, NSD2, and NSD3 mRNA in HPV+ and HPV- HNSCC. In addition, overall survival outcomes were compared for both the HPV+ and HPV- subsets of patients based on stratification by NSD1, NSD2, and NSD3 expression levels. Results Expression levels of NSD1, NSD2 or NSD3 were not correlated with altered lymphocyte infiltration in HPV+ HNSCC. More importantly, low expression of NSD1, NSD2, or NSD3 correlated with significantly reduced overall patient survival in HPV+, but not HPV- HNSCC. Conclusion These results starkly illustrate the contrast in molecular features between HPV+ and HPV- HNSCC tumors and suggest that NSD1, NSD2, and NSD3 expression levels should be further investigated as novel clinical metrics for improved prognostication and patient stratification in HPV+ HNSCC.

scholarly journals The Functional Quality of Soluble Recombinant Polypeptides Produced in Escherichia coli Is Defined by a Wide Conformational Spectrum

2008 ◽  
Vol 74 (23) ◽  
pp. 7431-7433 ◽  
Author(s):  
Mónica Martínez-Alonso ◽  
Nuria González-Montalbán ◽  
Elena García-Fruitós ◽  
Antonio Villaverde
Keyword(s):  
Escherichia Coli ◽  
Green Fluorescent Protein ◽  
Recombinant Proteins ◽  
Fluorescent Protein ◽  
Native Structure ◽  
Functional Quality ◽  
Soluble Aggregates ◽  
Recombinant Polypeptides ◽  
Green Fluorescent

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.

scholarly journals Use of a Beet necrotic yellow vein virus RNA-5-derived replicon as a new tool for gene expression

2005 ◽  
Vol 86 (2) ◽  
pp. 463-467 ◽  
Author(s):  
Laure Schmidlin ◽  
Didier Link ◽  
Jérôme Mutterer ◽  
Hubert Guilley ◽  
David Gilmer
Keyword(s):  
Gene Expression ◽  
Recombinant Proteins ◽  
Expression System ◽  
Green Fluorescence Protein ◽  
Yellow Vein ◽  
Expression Levels ◽  
Virus Rna ◽  
New Gene ◽  
Infected Cells ◽  
Fluorescence Protein

A new gene-expression system based on RNA-5 of Beet necrotic yellow vein virus (BNYVV) was constructed to allow the expression of recombinant proteins in virally infected cells. Replication and expression levels of the RNA-5-based replicon containing the green fluorescence protein (GFP) gene were compared with those obtained with the well-characterized RNA-3-derived replicon (Rep-3). When RNA-3 and/or RNA-4 BNYVV RNAs were added to the inoculum, the expression levels of RNA-5-encoded GFP were considerably reduced. To a lesser extent, RNA-3-derived GFP expression was also affected by the presence of RNA-4 and -5. Both RNA-3- and RNA-5-derived molecules were able to express proteins within the same infected cells. Together with Rep-3, the RNA-5-derived replicon thus provides a new tool for the co-expression of different recombinant proteins. In Beta macrocarpa, Rep-5-GFP was able to move in systemic tissues in the presence of RNA-3 and thus provides a new expression system that is not restricted to the inoculated leaves.

scholarly journals Production of recombinant proteins from Plasmodium falciparum in Escherichia coli

Biomédica ◽  
2016 ◽  
Vol 36 ◽  
Author(s):  
Ángela Patricia Guerra ◽  
Eliana Patricia Calvo ◽  
Moisés Wasserman ◽  
Jacqueline Chaparro-Olaya
Keyword(s):  
Escherichia Coli ◽  
Plasmodium Falciparum ◽  
Recombinant Proteins

<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>

scholarly journals Improving yield and quality of recombinant proteins in Escherichia coli by stress minimisation and mutant selection

2009 ◽  
Vol 25 ◽  
pp. S244
Author(s):  
Y. Sevastsyanovich ◽  
S. Alfasi ◽  
L. Griffiths ◽  
T. Overton ◽  
J. Cole
Keyword(s):  
Escherichia Coli ◽  
Recombinant Proteins ◽  
Mutant Selection ◽  
Yield And Quality

A Sugar-Inducible Excretion System for the Production of Recombinant Proteins with Escherichia coli

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 ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Proteins
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