![](images/graphics/blank.gif)
Recombinant soluble expression
-
Ebook "Heterologous gene expression in E. coli: Methods and protocols" herein describe methods, for example, to successfully express proteins in E. coli that would otherwise form aggregates in this host, to add post-translational modifications, to incorporate non-standard amino acid residues or moieties into E. coli expressed proteins, to identify binding partners, and to express membrane proteins.
312p
dongmelo
27-05-2024
6
2
Download
-
In this study, the DNA fragment encoding for the C-terminus domain of the AopB from Aeromonas hydrophila AH-1 was cloned into pET-M expression vector and expressed in Escherichia coli BL21 (DE3) host cells. The recombinant AopB-C-terminus domain was successfully purified using immobilized nickel affinity chromatography as a soluble form. Crosslinking analysis among AopB-C-terminus molecules in solution showed that this domain existed as a mixture of tetramer, trimer, dimer, and monomer forms.
6p
guitaracoustic02
08-12-2021
18
3
Download
-
In the present study, attempts were made to produce biologically active recombinant mouse hephaestin as a secretory form tagged with green fluorescent protein (GFP), Hpsec-GFP. Plasmid expressing Hpsec-GFP was constructed and transfected into COS and CHO cells. The GFP aided the monitoring expression in real time to select the best conditions to maximise expression and provided a tag for purifying and analysing Hpsec-GFP. The protein had detectable oxidase activity as shown by in-gel and solution-based assays.
13p
thiencuuchu
27-11-2021
8
1
Download
-
The recombinant miraculin was 19.97 ng per µg of the total soluble protein and equivalently with approximately 2% of the total soluble protein. For the first time, a taste modifying miraculin was successfully expressed in tobacco hairy root. The results in this study have given a promising approach for the application of the transgenic hairy root system to produce recombinant miraculin.
10p
spiritedaway36
25-11-2021
10
1
Download
-
Over the last 20 years in biotechnology, the production of recombinant proteins has been a crucial bioprocess in both biopharmaceutical and research arena in terms of human health, scientific impact and economic volume. Although logical strategies of genetic engineering have been established, protein overexpression is still an art.
16p
vikentucky2711
26-11-2020
7
1
Download
-
Porcine epidemic diarrhea virus (PEDV) is a highly contagious enteric pathogen of swine. The spike glycoprotein (S) of PEDV is the major immunogenic determinant that plays a pivotal role in the induction of neutralizing antibodies against PEDV, which therefore is an ideal target for the development of subunit vaccine.
9p
viuchiha2711
21-04-2020
15
1
Download
-
Recombinant proteins of therapeutic use are ideally produced in human cells to ensure appropriate co- and post-translational modifications. However, purification of secreted proteins from the culture media is impeded by low expression from transfected cell lines and the presence of serum proteins.
11p
viuchiha2711
21-04-2020
15
0
Download
-
Neurotrophic factors influence survival, differentiation, proliferation and death of neuronal cells within the central nervous system. Human ciliary neurotrophic factor (hCNTF) has neuroprotective properties and is also known to influence energy balance.
11p
viuchiha2711
21-04-2020
8
0
Download
-
Human tissue plasminogen activator (tPA) belongs to the serine protease family. It converts plasminogen into plasmin and is used clinically to treat thrombosis. Human tPA is composed of 527 amino acids residues and contains 17 disulfide bonds. Escherichia coli has been used only rarely for the efficient production of recombinant tPA.
9p
viuchiha2711
21-04-2020
13
0
Download
-
Manganese peroxidase (MnP) from Irpex lacteus F17 has been shown to have a strong ability to degrade recalcitrant aromatic pollutants. In this study, a recombinant MnP from I. lacteus F17 was expressed in Escherichia coli Rosetta (DE3) in the form of inclusion bodies, which were refolded to achieve an active enzyme.
15p
vihamax2711
21-04-2020
9
0
Download
-
Recombinant protein purification is a crucial step for biochemistry and structural biology fields. Rapid robust purification methods utilize various peptide or protein tags fused to the target protein for affinity purification using corresponding matrices and to enhance solubility.
11p
vihamax2711
21-04-2020
33
0
Download
-
Recombinant production of amebic cysteine proteases using Escherichia coli cells as the bacterial system has become a challenging effort, with protein insolubility being the most common issue.
7p
vihamax2711
21-04-2020
11
0
Download
-
Anti-NMDA receptor encephalitis (ANRE) is a potentially lethal disease attributed to auto-antibodies against the N-methyl-D-aspartate receptor (NMDAR). Full recovery is possible if therapy is initiated early in the disease course. Detection of ANRE antibodies in the cerebrospinal fluid (CSF) is essential for diagnosis.
9p
vihamax2711
21-04-2020
14
0
Download
-
Protein solubility characteristics are important determinants of success for recombinant proteins in relation to expression, purification, storage and administration. Escherichia coli offers a cost-efficient expression system.
10p
vihamax2711
21-04-2020
12
1
Download
-
Expression of microbial target genes in Escherichia coli is broadly used due to its advantages namely: well established system, easy to manipulate, a huge biomass, high level productivity, safe and inexpensive to grow. Metagenomic technique has been applying in Vietnam recently for effective mining of uncultured gene resources, especially in endemic mini-ecologies such as hot springs where the cell densities are low. DNA metagenome of Binh Chau hot spring was isolated and sequenced by Illumia HiseqTM.
8p
gaunguyen6789
22-10-2019
18
1
Download
-
The technology of recombinant single chain variable fragments (scFvs) expression has been used in research, diagnosis and treatment of diseases. In the previous study, we studied the expression of a recombinant single chain variable fragment recognizing blood A antigen (antiA-scFv) in E. coli. However, the protein was insoluble form resulting in difficulty for purification, refolding and activity assesment. Here, we present the study on fused expression of the recombinant scFv - specific blood A antigen with thioredoxin (Trx) in the expression vector pET32a(+).
8p
gaunguyen6789
22-10-2019
10
1
Download
-
The recombinant vector was screened by PCR method, sequenced and aligned with designed sequences. The in-frame vector was then introduced into E. coli BL21(DE3) for expression by inducing with 0.5mM IPTG. Fusion protein was purified using Ni-affinity chromatography. The results showed that the fused genes were in-frame cloned and completely matched with the designed sequences. SDS-PAGE and Western blot analyses showed Co1-GFP protein expressed in soluble form and could be purified at one-step elution of 500mM imidazole.
6p
nguyenhong1235
28-11-2018
29
0
Download
-
Study on the impact of temperature on the gene expression showed that TrxFljB was synthesized at lower level at 37oC comparing to the levels at 22oC and 25oC. 13% of the protein synthesized at 37oC was inclusion body. Lower temperatures used during induction phase increased the solubility of the recombinant protein. About 97% of TrxFljB synthesized at 25oC was soluble.
9p
jangni2
19-04-2018
35
2
Download
-
Cytochrome P450foxy (P450foxy, CYP505) is a fused protein of cytochrome P450 (P450) and its reductase isolated from the fungus Fusarium oxysporum, which catalyzes the subterminal (x-1x-3) hydroxylation of fatty acids. Here, we produced, purified and characterized a fused recombinant protein (rP450foxy) using the Escherichia coli expression system. Purified rP450foxy was catalytically and spectrally indistinguishable from the native protein, but most of the rP450foxy was recovered in the soluble fraction of E. coli cells unlike the membrane-bound native protein....
8p
research12
01-06-2013
54
3
Download
-
A cDNA encoding 6-phosphofructo-2-kinase/fructose-2,6bisphosphatase was isolated from a Spinacia oleracea leaf library and used to express a recombinant enzyme in Escherichia coli and Spodoptera frugiperda cells. The insoluble protein expressed in E. coli was purified and used to raise antibodies. Western blot analysis of a protein extract from spinach leaf showed a single band of 90.8 kDa. Soluble protein was purified to homogeneity from S. frugiperda cells infected with recombinant baculovirus harboring the isolated cDNA. ...
11p
research12
01-06-2013
40
4
Download
CHỦ ĐỀ BẠN MUỐN TÌM
![](images/graphics/blank.gif)