Expression and Purification of HCV Core and Core-E1E2 Proteins in Different Bacterial Strains

Mirnurollahi, Seyyedeh Masumeh; Bolhassani, Azam; Irani, Shiva; Davoudi, Noushin

doi:10.15171/ijb.1249

Cited by 6 publications

(3 citation statements)

References 31 publications

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“…11 The encoding region of HCV core does not show high variation rate; where nucleotides and the corresponding amino acids sequence, among various strains of HCV, indicate a homology of 81-88% and 96%, respectively. 16 The core protein is a much-conserved basic protein, which makes the chief element of HCV nucleocapsid. Core protein has multiple tasks including structural protein of HCV nucleocapsid, influencing lipid metabolism, apoptosis, and immune response.…”

Section: Discussionmentioning

confidence: 99%

Effect of RNA Interference on the Hepatitis C Virus Core Expression in HepG2/C3A Cells Using Genotype 4 Isolates from Saudi Patients

Shier

Ali

El-Wetidy

et al. 2019

J Coll Physicians Surg Pak

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Section: Discussionmentioning

confidence: 99%

Effect of RNA Interference on the Hepatitis C Virus Core Expression in HepG2/C3A Cells Using Genotype 4 Isolates from Saudi Patients

Shier

Ali

El-Wetidy

et al. 2019

J Coll Physicians Surg Pak

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“…Expression and purification of the HCV core protein in E. coli by reverse staining method The BL21 (DE3) bacterial strain harboring pET-core (Accession number: AB047640, [17]) was inoculated in 100 ml TY2X medium and grown to an optical density of 0.7-0.8 at 600 nm. The expression of core protein was induced by adding 1 mM IPTG and the pellet was harvested at 3 h post-induction.…”

Section: Methodsmentioning

confidence: 99%

Different protein expression systems can influence the direction of the immune responses against HCV core protein in animal model

Mirnurollahi

Irani

Davoudi

et al. 2015

vacres

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Introduction: Hepatitis C virus (HCV) infection is a major public health problem which influences about 170 million people worldwide. Different types of vaccines have been designed using HCV structural proteins to control the viral infection. The core nucleocapsid protein is one of the most conserved proteins and a desirable target for HCV vaccines, especially the protein-based vaccines. In current study, we generated the core protein recombinantly in prokaryotic (Escherichia coli) and eukaryotic (Leishmania tarentolae) expression systems and compared the humoral immune responses stimulated by each protein in a BALB/c mice model. Methods: The expression of HCV core protein was performed using the prokaryotic pET-28a/ BL21 expression system and also the eukaryotic LEXSY expression system. The recombinant core proteins expressed in E. coli and Leishmania were purified using reverse staining method and affinity chromatography under native conditions, respectively. The purified core proteins were detected by SDS-PAGE and Western blotting using anti-His antibody and were assessed by NanoDrop spectrophotometer. Finally, the abilities of both recombinant core proteins to induce the effective humoral immune responses were evaluated using indirect ELISA. Results: Our data indicated a clear band of ~ 21 kDa for the purified HCV core proteins in both expression systems, confirmed by SDS-PAGE and Western blotting. The mice immunization with both recombinant core proteins was able to produce high levels of antibody isotypes (IgG1 and IgG2a) in comparison with the controls (p < 0.05). In addition, the level of IgG2a response was significantly higher in the group immunized with the core protein purified from leishmania compared to the protein generated from E. coli (p < 0.05). Conclusion: The recombinant core protein generated by the leishmania expression system could induce a Th1-biased immune response with respect to the increase of IgG2a to IgG1 ratio.

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“…Their structure, function, expression, value, shape, and presence are important (5)(6)(7)(8)(9)(10). Generally speaking, protein accounts for about 18% of the total mass of the human body (11)(12). The most important thing is that it is related to life phenomena.…”

Section: Introductionmentioning

confidence: 99%

Protein molecular defect detection method based on a neural network algorithm

Zheng

Kahrizi²

2020

Cell Mol Biol (Noisy-le-grand)

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Proteins, as the largest macromolecules in the body, are among the most important components of the body and play very vital and important roles.These substances are made up of a series of amino acid chains that, depending on the type of protein, the number of these amino acids can reach several thousand.Proteins function differently depending on the type and location of their presence, including enzymatic activity to catalyze the process, identify microbes and cancer cells, transport substances such as respiratory gases, and signalize. In the biochemical experiment, the problem of optimizing the detection of protein molecular defects, because of the randomness of the information, parameters, selection and setting, limits the detection accuracy of protein molecular defects. Based on the characteristics of fast learning speed and a robust network of neural network algorithm, a protein molecular defect detection method based on a neural network algorithm was proposed. Firstly, the protein secondary structure was predicted by the method of protein secondary structure prediction based on the generalized regression neural network to obtain the protein structural features; secondly, the protein defective molecular sequence classification model based on the neural network was used to classify the protein defective molecular sequence to achieve the protein molecular defect detection. The results showed that the detection accuracy of the proposed method was very high, which meets the needs of protein molecular defect detection, and has some application advantages compared with similar detection methods.

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Expression and Purification of HCV Core and Core-E1E2 Proteins in Different Bacterial Strains

Cited by 6 publications

References 31 publications

Effect of RNA Interference on the Hepatitis C Virus Core Expression in HepG2/C3A Cells Using Genotype 4 Isolates from Saudi Patients

Effect of RNA Interference on the Hepatitis C Virus Core Expression in HepG2/C3A Cells Using Genotype 4 Isolates from Saudi Patients

Different protein expression systems can influence the direction of the immune responses against HCV core protein in animal model

Protein molecular defect detection method based on a neural network algorithm

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