scholarly journals Improving the Recombinant Protein Expression of Human Galectin-3 in BL21 Bacterial Host System

2020 ◽  
pp. 111-115
Author(s):  
Praveen Kumar Vemuri ◽  
Nikhil Reddy Varakala ◽  
Divyanshu Dhakate ◽  
Tripura Ravavarapu ◽  
Faina Philberta Dumpala ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Bacterial Culture ◽  
Recombinant Protein Expression ◽  
Shuttle Vector ◽  
Bacterial Host ◽  
E Coli ◽  
His Tag ◽  
Sds Page ◽  
Galectin 3 ◽  
Characterization Studies

Background: Regardless of the broad explore in the territory of glycobiology concerning structure and capacity of glycans, lectins and glycosylation forms, numerous viewpoints are still left unexplored. Aim: In this study, we analyzed the effect of shuttle vector on the secretion of human galectin recombinant protein. Methods: The galectin was expressed in E. coli BL21 by growing the bacterial culture in SOC medium and purified by nickel-based affinity chromatography due to its His-tag. Results: After cell lysis the protein was identified as a single 29 KDa band by 12% SDS-PAGE. Conclusion: Characterization studies clearly revealed that the purified protein was indeed galectin 3.

scholarly journals Effect of Ethanol and IPTG on the Recombinant Jembrana Trans-Activator of Transcriptation Protein Expression

2018 ◽  
Vol 10 (3) ◽  
pp. 559-564
Author(s):  
Indriawati Indriawati ◽  
Mega Salfia ◽  
R Susanti ◽  
Endang Tri Margawati
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Experimental Research ◽  
Quantitative Data ◽  
Recombinant Protein Expression ◽  
E Coli ◽  
Qualitative And Quantitative ◽  
Vaccine Product ◽  
Sds Page ◽  
Preliminary Study

Jembrana diseases are caused by Jembrana Diseases Virus (JDV). The previous study showed that Jembrana Trans-Activator of Trancriptation (JTAT) recombinant protein is effective as a vaccine for Jembrana diseases. The production of JTAT protein needs to be optimized to obtain a higher amount of vaccine. High expression of JTAT protein will produce a high vaccine product. This study aimed to examine the effect of the addition of ethanol and IPTG in E. coli media on the expression of JTAT recombinant protein. This research was experimental research with factorial RAL design with a variation factor of ethanol and IPTG. Qualitatively, the induction of each IPTG, ethanol and interaction between the two could induce the expression of JTAT protein and could be identified with SDS-PAGE at ±11.8 kDa. Statistically, the induction of IPTG, ethanol and interaction between the two were not significantly different. Qualitative and quantitative data show that ethanol can induce JTAT protein expression. This result can be used as a preliminary study to test the effectiveness of ethanol as a substitute for IPTG in inducing the recombinant protein expression.

scholarly journals pTSara-NatB, an improved N-terminal acetylation system for recombinant protein expression in E. coli

PLoS ONE ◽  
2018 ◽  
Vol 13 (7) ◽  
pp. e0198715 ◽  
Author(s):  
Matteo Rovere ◽  
Alex Edward Powers ◽  
Dushyant Shailesh Patel ◽  
Tim Bartels
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Recombinant Protein Expression ◽  
E Coli

Effect of supplementing Moringa oleifera essential oils on milk quality and fatty acid profile in dairy sheep

2019 ◽  
Author(s):  
N. Hemamalini ◽  
S. Ezhilmathi ◽  
A. Angela Mercy
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Cell Biology ◽  
Genetic Manipulation ◽  
Recombinant Protein Expression ◽  
Coli Strain ◽  
Expression Vectors ◽  
Functional Domain ◽  
Post Translational Modifications ◽  
E Coli

Escherichia coli is the most extensively used organism in recombinant protein production. It has several advantages including a very short life cycle, ease of genetic manipulation and the well-known cell biology etc. which makes E. coli as the perfect host for recombinant protein expression. Despite many advantages, E. coli also have few disadvantages such as coupled transcription and translation and lack of eukaryotic post-translational modifications. These challenges can be overcome by adopting several strategies such as, using different E. coli expression vectors, changing the gene sequence without altering the functional domain, modified E. coli strain usage, changing the culture parameters and co-expression with a molecular chaperone. In this review, we present the level of strategies used to enhance the recombinant protein expression and its stability in E. coli.

Monitoring of transcriptome and proteome profiles to investigate the cellular response of E. coli towards recombinant protein expression under defined chemostat conditions

2008 ◽  
Vol 135 (1) ◽  
pp. 34-44 ◽  
Author(s):  
Karin Dürrschmid ◽  
Helga Reischer ◽  
Wolfgang Schmidt-Heck ◽  
Thomas Hrebicek ◽  
Reinhard Guthke ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Cellular Response ◽  
Recombinant Protein Expression ◽  
E Coli

scholarly journals Purification and reactivation of recombinant Synechococcus phytoene desaturase from an overexpressing strain of Escherichia coli

1993 ◽  
Vol 291 (3) ◽  
pp. 687-692 ◽  
Author(s):  
P D Fraser ◽  
H Linden ◽  
G Sandmann
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Deae Cellulose ◽  
Cellular Protein ◽  
Phytoene Desaturase ◽  
E Coli ◽  
Sds Page ◽  
Purification Scheme ◽  
Overexpressing Strain ◽  
Cellulose Column

The Synechococcus phytoene desaturase has been isolated from an overexpressing strain of Escherichia coli. The plasma pPDSde135 mediated the overexpression of the full-length polypeptide directly. The recombinant protein comprised 5% of the total cellular protein and was found predominantly in the inclusion body fraction. Urea was used to solubilize the recombinant protein from the inclusion fraction and the protein was subsequently purified to homogeneity on a DEAE-cellulose column. The purification scheme yielded 4.0 mg of homogeneous desaturase protein after a 20-fold purification, recovering 40% of the original protein from a 100 ml suspension culture of E. coli. The recombinant desaturase had an apparent molecular mass of 53 kDa on SDS/PAGE and crossreacted with an antiserum raised against the expressed protein. Desaturase activity was restored upon the removal of urea. The enzyme catalysed the conversion of phytoene to zeta-carotene via phytofluene. These products of the desaturase reaction existed predominantly in a cis configuration. Lipid replenishment enhanced activity. NAD+ and NADP+ were observed to be involved, whilst FAD was an ineffective electron acceptor.

scholarly journals Analytical and preparative biosynthesis of S100B recombinant protein using the pBT7-N-His vector system and preliminary studies of structure and function of this protein

2018 ◽  
pp. 161-164
Author(s):  
О.Л. Терёхина ◽  
М.К. Нурбеков ◽  
О.П. Дмитренко ◽  
Д.М. Давыдов
Keyword(s):  
Recombinant Protein ◽  
Molecular Mechanisms ◽  
The Body ◽  
S100b Protein ◽  
Vector System ◽  
Diagnostic Systems ◽  
E Coli ◽  
Sds Page ◽  
Bacterial Clone ◽  
And Function

С целью исследований структуры и функций белка S100B в клетке и в тканях был проведен цикл работ по оптимизации экспрессии рекомбинантного белка (рекS100B) в E. coli. Проведены процедуры аналитической экспрессии рекS100B в составе рекомбинантной плазмиды pBT7-N-His-S100B03. При SDS-ПААГЭ лизатов клонов бактерий выявлена четко экспрессирующаяся полоса в 10 кДа, которая была идентифицирована как мономерная форма белка. Перспективы исследований рекS100B связаны с потенциальным его использованием для изучения тонких молекулярных механизмов PPI взаимодействий в системе S100B/RAGE рецептор как ключевого звена передачи сигналов в клетке и организме и в качестве перспективного объекта создания диагностических систем мониторинга состояний организма в норме и при патологии связанной с нарушениями регуляции гена и/или функций S100B белка. To study structure and functions of the S100B protein in cells and tissues, a series of studies was conducted to optimize the recombinant protein (recS100B) expression in E. coli. Procedures for analytical expression of recS100B in the pBT7-N-His-S100B03 recombinant plasmid were performed. In SDS-PAGE of bacterial clone lysate, a clear 10 kDa band expression was detected, which was identified as a monomeric form of the protein. Prospects for the S100B study are related with its potential use for investigating molecular mechanisms of PPI interactions in the S100B/RAGE system as a key signal transducer in the cell and body and as a promising object for developing diagnostic systems for monitoring the body state in normal and pathological conditions associated with impaired regulation of the gene and/or functions of the S100B protein.

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