Cloning, expression, purification and antigenicity of antigenic region of MOMP protein from chlamydia trachomatis

Introduction: Chlamydia trachomatis is an obligate intracellular parasites, gram-negative bacteria and also is one of the most common sexually transmitted infections. Correspondingly, the MOMP protein contains more than 60 of the extra-membrane proteins, which indicates the importance of this protein for the detection of Chlamydia trachomatis. The purpose of the study is to produce recombinant protein composed of antigenic regions of the MOMP protein and its antigenicity. Materials and Methods: In this study, the regions with the highest antigenic property of the omp1 gene, with a length of 674 bp, were obtained based on bioinformatics software's and linked with flexible linker. Then, it was cloned and expressed in plasmid vector pET32a. After purification of the recombinant protein, its antigenicity was evaluated using Western blot technique with serum of infected people with Chlamydia. Results: Based on the results, in all serum samples of patients with chlamydial infections, antibody response bands were observed in nitrocellulose paper. Anyway, no antibody response band was detected in the serum samples of healthy people. Conclusion: Data showed that antigenic region of MOMP protein can be expressed by in E. coli. This protein was recognized by sera patients suffering from Chlamydia trachomatis infection. Conclusively, the recombinant protein has similar epitopes and close antigenic properties to the natural form of this antigen. © 2017, Semnan University of Medical Sciences. All rights reserved

Cloning, expression, purification and antigenicity of antigenic region of MOMP protein from chlamydia trachomatis

Introduction: Chlamydia trachomatis is an obligate intracellular parasites, gram-negative bacteria and also is one of the most common sexually transmitted infections. Correspondingly, the MOMP protein contains more than 60 of the extra-membrane proteins, which indicates the importance of this protein for the detection of Chlamydia trachomatis. The purpose of the study is to produce recombinant protein composed of antigenic regions of the MOMP protein and its antigenicity. Materials and Methods: In this study, the regions with the highest antigenic property of the omp1 gene, with a length of 674 bp, were obtained based on bioinformatics software's and linked with flexible linker. Then, it was cloned and expressed in plasmid vector pET32a. After purification of the recombinant protein, its antigenicity was evaluated using Western blot technique with serum of infected people with Chlamydia. Results: Based on the results, in all serum samples of patients with chlamydial infections, antibody response bands were observed in nitrocellulose paper. Anyway, no antibody response band was detected in the serum samples of healthy people. Conclusion: Data showed that antigenic region of MOMP protein can be expressed by in E. coli. This protein was recognized by sera patients suffering from Chlamydia trachomatis infection. Conclusively, the recombinant protein has similar epitopes and close antigenic properties to the natural form of this antigen. © 2017, Semnan University of Medical Sciences. All rights reserved

Cloning, expression and purification of escherichia coli modified phytase

Introduction: Phytases are the class of phosphatases, which are capable of hydrolyzing phytic acid. Phytases with the phytate degradation are able to reduce or eliminate the harmful effects of phytate. Acidic and thermal stable phytase with high yield and purity by a relatively inexpensive system had extensive application. So, in this study, by modification in enzyme sequence, recombinant phytase production with the shorter length and high expression level was assessed. Materials and Methods: The phytase gene sequence was obtained from the NCBI database. After bioinformatics studies and doing the noted modification for increasing protein expression, gene proliferation was done by using PCR. E. coli BL21 (DE3) was used to express the protein. Protein purification was performed by Ni-NTA kit and finally, enzyme activity was assessed. Results: Phytase was successfully expressed and purified. Enzyme activity assay showed a significant activity. Conclusion: Produced recombinant phytase had high activity in spite of eliminating parts of the enzyme. © 2017, Semnan University of Medical Sciences. All rights reserved

Epitope Prediction by Novel Immunoinformatics Approach: A State-of-the-art Review

Immunoinformatics is a science that helps to create significant immunological information using bioinformatics softwares and applications. One of the most important applications of immunoinformatics is the prediction of a variety of specific epitopes for B cell recognition and T cell through MHC class I and II molecules. This method reduces costs and time compared to laboratory tests. In this state-of-the-art review, we review about 50 papers to find the latest and most used immunoinformatic tools as well as their applications for predicting the viral, bacterial and tumoral structural and linear epitopes of B and T cells. In the clinic, the main application of prediction of epitopes is for designing peptide-based vaccines. Peptide-based vaccines are a considerably potential alternative to low-cost vaccines that may reduce the risks related to the production of common vaccines. © 2019, Springer Nature B.V

The attentive focus on T cell-mediated autoimmune pathogenesis of psoriasis, lichen planus and vitiligo

T cell-mediated autoimmune skin diseases develop as a result of the aberrant immune response to the skin cells with T cells playing a central role. These chronic inflammatory skin diseases encompass various types including psoriasis, lichen planus and vitiligo. These diseases show similarities in their immune-pathophysiology. In the last decade, immunomodulating agents have been very successful in the management of these diseases thanks to a better understanding of the pathophysiology. In this review, we will discuss the immunopathogenic mechanisms and highlight the role of T lymphocytes in psoriasis, lichen planus and vitiligo. This study could provide new insights into a better understanding of targeted therapeutic pathways and biological therapies. © 2020 The Scandinavian Foundation for Immunolog

Saw-Cutting Guidelines for Concrete Pavements: Examining the Requirements for Time and Depth of Saw-Cutting

Joints are placed in Portland cement concrete pavements (PCCP) to control random cracking. These joints provide a weakened plane that enables a crack to form in a controlled manner, relieving residual stresses that develop when thermal, hygral, or hydration movements are resisted by sub grade and adjoining pavement. While the concept of creating a weakened plane through saw-cutting is straightforward, determining the time and depth of the saw-cut has proven to be complicated. The goal of this project was to reduce the risk for joint raveling and random cracking. Specifically, this project has focused on: developing a procedure for determining the appropriate saw-cutting time window for typical pavements constructed in the state of Indiana, determining the depth of the saw-cut that minimizes the risk of micro-cracking and random crack development, and developing tools and training materials for paving contractors and state inspectors that aid in implementing the findings of this study in concrete pavements. Toward this end the project was divided into three phases. The first phase of the project consisted of shadowing five pavement projects in Indiana. From these field investigations practical information was gained which was useful in developing the laboratory testing program. The second phase of this work involved in the development of the laboratory testing program. This phase in large part was involved in the development and commissioning of a new tensile wedge testing approach to determine the early age properties of concrete. Finally, finite element simulations were performed to simulate the early-age behavior of pavements constructed under a variety of saw-cutting sequences, environmental conditions. A strength reduction factor was computed based on the depth of the saw-cut. It was shown that the time of the saw-cut introduction needs to occur before the residual stress divided by the product of the strength reduction factor and tensile strength was equal to unity. It was also shown that shallower saw-cut depths were more prone to construction and material property variability. Recommendations are made to assist contractors in determining when saw-cuts are placed that can greatly improve field operations

Predicting candidate epitopes on Ebolaviruse for possible vaccine development

Zaire ebolavirus a member of family Filoviridae is the cause of hemorrhagic fever. Due to lack of appropriate anti-viral or vaccine, this disease is very lethal. In this study we tried to find epitopes for superficial glycoprotein of Zaire ebolavirus (that have high antigenicity for MHC I, II and B cells) with use of in-silico methods and immunoinformatics approach. By use of CTLPred, SYFPEITHI and Propred web applications for MHC class I and SYFPEITHI and Propred1web applications for MHC class II we had been able to find epitopes (peptides) that have highest score. Also ElliPro, IgPred and Discotope web tools had been performed to predict B cells epitopes. The sequence "SRFTPQFLL" and "IFFLYDRLAS" were selected to be epitopes for MHCs molecules. The region 255 to 310 was selected for B cell epitope. It was expected these peptides could be stimulated immune response and used for designing multi-peptide vaccine against ZEV but these results should be reliable with experimental analysis. © 2015 ACM

Cloning and High-Level Expression of the Enzymatic Region of Phytase in E. coli

Phytase is an important enzyme poses great nutritional significance in humans and monogastric animals diets. The phytase production yield using wild sources, including micro-organisms, plants, and animals is sorely low. Thus, recombinant expression of phytase has received increasing interest for achieving production rate. Escherichia coli is the most preferred host for expression of heterologous proteins but overexpression of recombinant phytase in E. coli, met with limited success due to the sequestration of the enzyme into inclusion bodies. In the present study, artificial phytases gene with excellent thermostability and activity were designed by detecting the enzymatic region of the E. coli phytase gene by employing bioinformatics tools. Then, the PCR amplified recombinant gene was expressed in E. coli and the active enzyme was recovered from inclusion bodies. Employing cysteine amino acid in the dialysis buffer succeed to the superior activity of the enzyme with a specific activity of 73.8 U/mg. The optimum temperature and pH for enzyme activity were determined at 60 °C and 4, respectively. The novel recombinant enzyme illustrated perfect thermostability up to 70 °C with maintenance 75 of its activity. The enzyme was stable at pH range of 2�10. Moreover, the effects of ions and chemical compounds on enzyme stability and activity were assessed. © 2019, Springer Nature B.V