scholarly journals Expression of Hepatitis B viral surface antigen (Pre S1) in E. coli

2020 ◽  
Vol 11 (3) ◽  
pp. 3046-3052
Author(s):  
Shahad Basel Ismail ◽  
Yaseen Ismael Mamoori ◽  
Ibrahim Abdulla Ahmed
Keyword(s):  
Molecular Weight ◽  
Hepatitis B ◽  
Protein Band ◽  
Effective Vaccine ◽  
Chronic Hepatitis B Infection ◽  
Colony Pcr ◽  
E Coli ◽  
Sds Page ◽  
B Virus ◽  
Viral Surface

Hepatitis B is the most common liver diseases, which caused by hepatitis B virus (HBV) infection. There are around 257 million people around the world suffer from severe chronic hepatitis B infection. Therefore, it is necessary to develop a vaccine to prevent viral infection. PreS1 is one of the HBV envelope proteins that have been proved to be an effective vaccine. Accordingly, Viral DNA was purified from patients’ serums and amplified by PCR using specific primers. Amplicons of 324 bp bands of PreS1 was observed on gel electrophoresis. The PreS1 was cloned into pTXB21 plasmid to form the recombinant plasmid pTXB1_PreS1 and transformed into DH5α E. coli. Screening of transformants was done using Colony PCR and Sequencing. Alignment of 26 polypeptide sequences showed conservation of this region. The pTXB1_PreS1 was retransformed into T7 Express Competent E. coli and screened using colony PCR. The PreS1 was expressed as a recombinant protein fused to an intein tag with a molecular weight of ~ 39.5 kD. The PreS1 protein was purified by a single affinity chromatography step and after cleaved from intein tag by Dithiothreitol the obtained protein had a molecular weight of ~ 11.5 kD. Only one protein band was observed on the SDS-page gel. The PreS1 protein was successfully cloned and expressed in E. coli, which can be used as a vaccine against HBV.

Erratum to “Characterization and diagnostic potential of hepatitis B virus nucleocapsid expressed in E. coli and P. pastoris”

2002 ◽  
Vol 102 (1-2) ◽  
pp. 173
Author(s):  
Bénédicte Watelet ◽  
Martine Quibriac ◽  
Dominique Rolland ◽  
Gaspard Gervasi ◽  
Marie Gauthier ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
E Coli ◽  
Diagnostic Potential ◽  
B Virus

Erratum to “Characterization and diagnostic potential of hepatitis B virus nucleocapsid expressed in E. coli and P. pastoris”

2002 ◽  
Vol 102 (1-2) ◽  
pp. 175-190 ◽  
Author(s):  
Bénédicte Watelet ◽  
Martine Quibriac ◽  
Dominique Rolland ◽  
Gaspard Gervasi ◽  
Marie Gauthier ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
E Coli ◽  
Diagnostic Potential ◽  
B Virus

PROTEIN PROFILING OF DIFFERENT PLANT TISSUES FROM HERB PHYLLANTHUS NIRURI

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Ainul Mardhiah Mohd Nail ◽  
Noor Hasniza Md Zin
Keyword(s):  
Molecular Weight ◽  
Protein Band ◽  
Protein Profiling ◽  
Protein Extract ◽  
Phyllanthus Niruri ◽  
Two Dimensional Gel Electrophoresis ◽  
Plant Parts ◽  
Sds Page ◽  
Isoelectric Points

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anticancer, antibacterial, antioxidant, anti-hypertensive and also anti-diabetic properties.  In this research, the proteomic part of P. niruri was studied to determine the bioactive peptides that responsible for specific characteristics. Total soluble proteins from different plant parts of freshly collected P. niruri were extracted using TCA/acetone method and then quantified using Bradford assay. Fruits part was found to have a significantly higher amount of proteins (4.91µg/µl + 0.21) compared to leaves (4.18µg/µl + 0.15). To determine the quality of proteins in the crude extract, SDS-Page was carried out which separates proteins in the basis of molecular weight. Proteins extracted from leaves were widely distributed between the range of 3.5 kDa to 160 kDA. Meanwhile, proteins in fruits mainly distributed within the range of 15 kDa to 80 kDa. The most highly expressed protein band was found in fruit, located in between 30 to 40 kDa. The protein extracts were then further analyzed based on the molecular weight and isoelectric points using two-dimensional gel electrophoresis (2D-GE) approach. Based on the profile pattern obtained from 2D-GE analysis, protein extract from fruits seems to express more protein spots compared to protein extract from leaves. Protein spots from fruit are seen to be intensely resolved within pH 4 to 10 at molecular weight between 10 kDa to 80 kDa. On the other hand, protein spots from leaves were moderately resolved at pH 4 to 10 at molecular weight within 10 kDa to 50 kDa.

scholarly journals Állásfoglalás az egészségügyi dolgozók hepatitis B-vírus-fertőzéssel szembeni immunizációjáról

2019 ◽  
Vol 160 (41) ◽  
pp. 1607-1616
Author(s):  
Csilla Jekkel ◽  
Beáta Onozó ◽  
Petra Scharek ◽  
Andrea Kulcsár
Keyword(s):  
Health Care ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Occupational Exposures ◽  
Hbv Infection ◽  
Effective Vaccine ◽  
Care Workers ◽  
B Virus ◽  
Increasing Risk ◽  
Induced Immunity

Abstract: More than 200 million HBV surface antigen (HBsAg) positive, hepatitis B virus (HBV) carriers live worldwide. Health-care personnel have increasing risk for aquiring the HBV infection. An effective vaccine is available against the infection, however, a certain proportion of the vaccinated patients do not respond to the vaccine depending on certain factors. Therefore, vaccine-induced immunity (anti-HBs) should be tested at health-care workers. For nonresponders, there are other vaccination strategies to try to achieve protection. This recommendation also provides a guidance for postexposure prophylaxis following occupational exposures against HBV infection. This is the first Hungarian recommendation about this topic. Orv Hetil. 2019; 160(41): 1607–1616.

scholarly journals In Vitro Systems for Studying Different Genotypes/Sub-Genotypes of Hepatitis B Virus: Strengths and Limitations

Viruses ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 353 ◽  
Author(s):  
Constance N. Wose Kinge ◽  
Nimisha H. Bhoola ◽  
Anna Kramvis
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
In Vitro Model ◽  
Antiviral Agents ◽  
Model Systems ◽  
Effective Vaccine ◽  
B Virus ◽  
In Vitro Systems ◽  
Vitro Model

Hepatitis B virus (HBV) infects the liver resulting in end stage liver disease, cirrhosis, and hepatocellular carcinoma. Despite an effective vaccine, HBV poses a serious health problem globally, accounting for 257 million chronic carriers. Unique features of HBV, including its narrow virus–host range and its hepatocyte tropism, have led to major challenges in the development of suitable in vivo and in vitro model systems to recapitulate the HBV replication cycle and to test various antiviral strategies. Moreover, HBV is classified into at least nine genotypes and 35 sub-genotypes with distinct geographical distributions and prevalence, which have different natural histories of infection, clinical manifestation, and response to current antiviral agents. Here, we review various in vitro systems used to study the molecular biology of the different (sub)genotypes of HBV and their response to antiviral agents, and we discuss their strengths and limitations. Despite the advances made, no system is ideal for pan-genotypic HBV research or drug development and therefore further improvement is required. It is necessary to establish a centralized repository of HBV-related generated materials, which are readily accessible to HBV researchers, with international collaboration toward advancement and development of in vitro model systems for testing new HBV antivirals to ensure their pan-genotypic and/or customized activity.

scholarly journals Purification and parcial characterization of a hemagglutinating factor (HAF): a possible adhesive factor of the diffuse adherent of Escherichia coli (DAEC)

1996 ◽  
Vol 38 (6) ◽  
pp. 401-406 ◽  
Author(s):  
Yano Tomomasa ◽  
Cleide Ferreira Catani ◽  
Michiko Arita ◽  
Takeshi Honda ◽  
Toshio Miwatani
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Hela Cells ◽  
Immunofluorescence Test ◽  
Native Form ◽  
Bacterial Surface ◽  
E Coli ◽  
Sds Page ◽  
Adhesive Factor

The mannose-resistant hemagglutinating factor (HAF) was extracted and purified from a diffuse adherent Escherichia coli (DAEC) strain belonging to the classic enteropathogenic E. coli (EPEC) serotype (0128). The molecular weight of HAF was estimated to be 18 KDa by SDS-PAGE and 66 KDa by Sephadex G100, suggesting that the native form of HAF consists of 3-4 monomeric HAF. Gold immunolabeling with specific HAF antiserum revealed that the HAF is not a rigid structure like fimbriae on the bacterial surface. The immunofluorescence test using purified HAF on HeLa cells, in addition to the fact that the HAF is distributed among serotypes of EPEC, suggests that HAF is a possible adhesive factor of DAEC strains

scholarly journals A Recombinant Multiepitope Protein for Hepatitis B Diagnosis

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Marilen Queiroz de Souza ◽  
Alexsandro Sobreira Galdino ◽  
José Carlos dos Santos ◽  
Marcus Vinicius Soares ◽  
Yanna C. de Nóbrega ◽  
...  
Keyword(s):  
Hepatitis B ◽  
Time Course ◽  
Clinical Stage ◽  
Acute Infection ◽  
Single Step ◽  
Liver Inflammation ◽  
E Coli ◽  
Terminal Time ◽  
Gene Coding ◽  
B Virus

Hepatitis B is a liver inflammation caused by hepatitis B virus (HBV) and can be diagnosed in clinical stage by hepatitis B core antibody from IgM class (anti-HBcIgM). Hepatitis B core antibody from IgG class (Anti-HBcIgG) appears quickly after IgM, reaching high titers in chronic hepatitis, and remains even after cure. Since hepatitis B core antibody (anti-HBc) is the first antibody identified and sometimes the only marker detected during the course of infection, it can be used both to indicate HBV acute infection (anti-HBc-IgM) and to identify individuals who have come into contact with the virus (anti-HBc-IgG). In this work we propose a recombinant hepatitis B core multiepitope antigen (rMEHB) to be used for diagnosis of hepatitis B. For this purpose, a synthetic gene coding for rMEHB was designed and cloned into vector pET21a with a 6xHis tag at the C-terminal. Time course induction inE. colishowed an induced protein with an apparent molecular mass of ~21 kDa. Protein purification was performed by a single step with affinity chromatography Ni-NTA. Circular dichroism spectroscopy indicated rMEHB as a thermal stable protein at pH 7.0 and 8.0. In these conditions rMEHB was successfully used to perform an enzyme linked immuno sorbent assay (ELISA) with positive and negative sera.

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