Optimization of induction conditions and purification of glycoprotein B cytomegalovirus
محل انتشار: بیستمین کنگره بین المللی میکروب شناسی ایران
سال انتشار: 1398
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 453
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شناسه ملی سند علمی:
MEDISM20_143
تاریخ نمایه سازی: 26 بهمن 1398
چکیده مقاله:
Introduction and Objectives: In recent years, cytomegalovirus has been identified as an important cause of neonatal congenital abnormalities, and important factors transmission infection including blood transition, blood products and organ transplants, are known. Some viral glycoproteins play an important role in the life cycle of the virus. Among them, glycoprotein B (gB) is a surface glycoprotein virus. gB is site for binding anti-virus antibodies that can be used to target subunit vaccines.Materials and Methods: In this study, to produce gB in Full Length, after bioinformatics and optimization studies, the sequence of this gene was synthesized in pET15b vector and then transformed into E. coli BL21. In order to obtain the gB protein, Ni-NTA chromatography resin was purified using microtubes method. Optimization of protein expression in prokaryotic system was performed by evaluating effective parameters and SDS PAGE and Western Blot were used to confirm the gene expression vector. An expressive protein was prepared for final purification and immunological evaluations. Results: The induction of transfected BL21 bacteria with IPTG resulted in expression of gB, The highest yield of recombinant gB occurred in the presence of 1 mM of IPTG in LB Broth medium after 8 h at 37°C. Thereby purifying the chromatography and purification of this protein and confirming the western blot. Conclusion: considering the ability to express the glycoprotein expressed in the BL21 bacteria, this protein can be a suitable candidate for immunological evaluation and vaccine production against cytomegalovirus.
کلیدواژه ها:
نویسندگان
Somayeh Azadfar
Department of Biology, Faculty of basic Biological Sciences, North-Tehran Branch, Islamic Azad University, Tehran, Iran.
Ahmad Majd
Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources
Ahad Yamchi
Infectious diseases research center, Golestan University of Medical Sciences, Gorgan, Iran
Hadi Razavi Nikoo
Department of Biology, Faculty of basic Biological Sciences, North-Tehran Branch, Islamic Azad University, Tehran, Iran.
Alijan Tabarraei
Department of Biology, Faculty of basic Biological Sciences, North-Tehran Branch, Islamic Azad University, Tehran, Iran.